Methods and Compositions for Treating Striae Distensae

ABSTRACT

The present invention relates generally to methods of use and compositions useful for treating, preventing, and or reducing the appearance of stretch marks. The composition includes a combination of vegetable amino acids,  Myrciaria dubia  fruit extract,  Peucedanum graveolens  extract, and tetrahexyldecyl ascorbate.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/403,912 filed Oct. 4, 2016, the content of which is incorporated intothe present application by reference.

BACKGROUND OF THE INVENTION A. Field of the Invention

The present invention relates generally to compositions that can be usedto improve the skin's condition and/or visual appearance. In certainaspects, the compositions of the present invention can include, forexample, a combination of ingredients to reduce the appearance of,prevent, and/or treat striae, commonly called stretch marks. Thiscombination of ingredients can be included in a wide-range of productformulations (e.g., serums, creams, cleansers, toners, gels, emulsions,gel emulsions, gel serums, etc.).

B. Description of Related Art

Striae or striae distensae, also known as stretch marks, are areas ofthe skin with the appearance of having bands, stripes, or lines thereon.Striae are most commonly located on a person's breasts, hips, thighs,buttocks, abdomen, and/or flank. These marks may appear when there is arapid stretching of the skin (e.g., during pregnancy, weight gain,building muscle mass, rapid growth such as seen with adolescence, etc.).This rapid stretching can cause collagen fibers just under the surfaceof the skin to become damaged, which can result in the appearance ofstretch marks on the skin surface. Stretch marks can often appear red,thinned, and/or glossy and may eventually become whitish and scar-likein appearance.

While stretch marks are undesired, current products on the market eitherdo not effectively address the treatment, prevention, and/or appearanceof striae and/or they have unwanted side effects such as causingmalformations in unborn children and/or causing skin irritation. SeeMedications and More During Pregnancy and Breastfeeding, Retinoids;California Department of Public Health. For example, current productsmay not address the visibility of stretch marks, the loss of skinfirmness, redness, uneven skin tone, decreased elasiticy, increasedroughness of the skin, and/or increased thickness of the stretch mark.Current products also may not address some of the underlying factorscausing stretch marks such as an overly oriented collagen network,decreased collagen density, and/or decreased fibrillin-1.

SUMMARY OF THE INVENTION

The inventors have identified a solution to the problems associated withtreating striae (the terms striae, striae distensae, and stretch markscan be used interchangeably throughout the specification). The solutionresides in a combination of ingredients including an aqueous extract ofnavy bean, which includes vegetable amino acids in the extract, anaqueous extract of Myrciaria dubia (camu camu) fruit pulp, an aqueousextract of Peucedanum graveolens (dill) extract, and tetrahexyldecylascorbate. This combination can be used to create topical skincompositions to treat/reduce the appearance of or prevent the formationof striae. This combination can also be used to create topical skincompositions that inhibit COX-1, MMP1, MMP3, MMP9, and/or tyrosinaseactivity, that induce collagen production, and/or that provide anantioxidant capacity. Specifically, and as illustrated in the Examples,this combination of ingredients has been shown to decrease visibility ofstretch marks, beneficially increase overall appearance of stretchmarks, decrease redness of stretch marks, increase smoothness of stretchmarks, increase skin tone/evenness of stretch marks, increase elasticityof stretch marks, increase firmness of stretch marks, improve texture ofstretch marks, decrease the thickness of stretch marks, and/or decreaseechogenicity of subject's stretch marks. Without wishing to be bound bytheory, it is believed that the combination of ingredients serve as aneffective treatment of striae in that the combination can decreasecollagen network orientation, increase collagen density of stretchmarks, and increase fibrillin-1 expression in stretch marks. Inaddition, for skin that does not include stretch marks, the combinationcan increase collagen density and fibrillin-1 expression, which can behelpful in treating a range of skin conditions such as fine lines orwrinkles, sagging or loose skin, skin that has lost its elasticity, etc.

In some aspects, there is disclosed a topical composition. In someaspects the topical composition includes any one of, any combination of,or all of vegetable amino acids, Myrciaria dubia (camu camu) fruitextract, Peucedanum graveolens (dill) extract, and/or tetrahexyldecylascorbate. In some aspects the vegetable amino acids are an aqueousextract of navy bean. In some aspects the Myrciaria dubia fruit extractis an aqueous extract that comprises the pulp of Myrciaria dubia fruit.In some aspects the Peucedanum graveolens extract is an aqueous extract.In some aspects the composition is capable of increasing collagendensity in skin and/or striae. In some aspects the composition iscapable of increasing fibrillin-1 expression in skin and/or striae. Insome aspects the composition is capable of decreasing collagen networkorientation. In some aspects the composition is capable of decreasingstretch mark visibility. In some aspects the composition is capable ofimproving the overall appearance of a stretch mark. In some aspects thecomposition is capable of decreasing redness in skin and/or striae. Insome aspects the composition is capable of increasing smoothness in skinand/or striae. In some aspects the composition is capable of increasingskin tone/evenness in skin and/or striae. In some aspects thecomposition is capable of increasing elasticity in skin and/or striae.In some aspects the composition is capable of increasing firmness ofskin and/or striae. In some aspects the composition is capable ofimproving texture of skin and/or striae. In some aspects the compositionis capable of decreasing thickness of a stretch mark. In some aspectsthe composition is capable of decreasing echogenicity of striae. In someaspects, the composition is capable of inhibiting COX-1, MMP1, MMP3,MMP9, and/or tyrosinase activity. In some aspects, the composition iscapable of increasing collagen production. In some aspects, thecomposition is capable of inhibiting oxidation of the skin and/orproviding an antioxidant to the skin and/or a formulation.

The amounts of the ingredients within the composition can vary (e.g.,amounts can be as low as 0.000001% to as high as 98% w/w or any rangetherein). In some aspects the composition includes 0.001 to 5% by weightof vegetable amino acids, 0.001 to 5% by weight of Myrciaria dubia fruitextract, 0.001 to 3% by weight of Peucedanum graveolens extract, and/or0.001 to 1% by weight of tetrahexyldecyl ascorbate. In some aspects, thecomposition includes a sufficient amount of a combination of vegetableamino acids, Myrciaria dubia fruit extract, Peucedanum graveolensextract, and tetrahexyldecyl ascorbate to treat striae. In some aspects,the composition includes a sufficient amount of Myrciaria dubia fruitextract to inhibit COX-1, MMP1, MMP3, MMP9, and/or tyrosinase activity,induce collagen production, and/or provide an antioxidant capacityand/or reduce oxidative damage to skin. The composition may furthercomprise one or more ingredients described herein. For example, thecomposition may comprise one or more additional ingredients selectedfrom one or more conditioning agents, moisturizing agents, pH adjusters,structuring agents, inorganic salts, and preservatives. In one aspect,the compositions disclosed herein are used to reduce the appearance of,prevent, and/or treat striae.

In some aspects, the topical composition further contains water. In someaspects the topical composition contains 60 to 85% by weight of water.In some aspects, the topical composition further contains pentyleneglycol, cyclopentasiloxane, glycerin, and hydroxyethyl acrylate/sodiumacryloyldimethyl taurate copolymer. In some aspects, the topicalcomposition contains 1 to 10% by weight of pentylene glycol, 1 to 5% byweight of cyclopentasiloxane, 0.1 to 5% by weight of glycerin, and 0.1to 5% by weight of hydroxyethyl acrylate/sodium acryloyldimethyl tauratecopolymer. In some aspects, the topical composition further containsdimethicone, dimethiconol, phenoxyethanol, and ethylhexylglycerin. Insome aspects, the topical composition contains PEG-12 dimethicone,triethanolamine, and disodium EDTA. In some aspects, the topicalcomposition contains 0.1 to 3% w/w of PEG-12 dimethicone, 0.1 to 3% w/wof triethanolamine, 0.01 to 1% w/w of disodium EDTA. In some aspects,the topical composition is an emulsion. In some aspects, the topicalcomposition is gel emulsion. In some aspects, the composition is aserum. In some aspects, the composition is a gel serum. In some aspects,the composition is a gel.

Methods of use for the compositions disclosed herein are also disclosed.In some aspects, a method is disclosed of improving a condition orappearance of skin, comprising applying any one of the compositionsdisclosed herein to skin in need thereof. In one aspect, any one of thecompositions disclosed herein are applied to skin and the composition isleft on the skin, or alternatively removed from the skin after a periodof time. In another aspect, the compositions disclosed herein are usedto treat and/or reduce the appearance of striae. In another aspect, thecompositions disclosed here are used to treat and/or reduce theappearance of striae by applying any one of the composition disclosedherein to striae, wherein the striae is treated and/or the appearance ofstriae is reduced. In another aspect, the compositions disclosed areused to prevent striae comprising applying any one of the compositiondisclosed herein to skin, wherein the striae are prevented.

In another aspect, a method for increasing collagen density of skinand/or a stretch mark is disclosed herein. In another aspect, a methodfor increasing fibrillin-1 expression of skin and/or a stretch mark isdisclosed herein. In another aspect, a method for decreasing collagennetwork orientation is disclosed herein. In another aspect, a method fordecreasing stretch mark visibility is disclosed herein. In anotheraspect, a method for improving the overall appearance of a stretch markis disclosed herein. In another aspect, a method for decreasing rednessof skin and/or a stretch mark is disclosed herein. In another aspect, amethod for increasing smoothness of skin and/or a stretch mark isdisclosed herein. In another aspect, a method for increasing skintone/evenness of skin and/or a stretch mark is disclosed herein. Inanother aspect, a method for increasing elasticity of skin and/or astretch mark is disclosed herein. In another aspect, a method forincreasing firmness of skin and/or a stretch mark is disclosed herein.In another aspect, a method for improving texture of skin and/or astretch mark is disclosed herein. In another aspect, a method fordecreasing thickness of a stretch mark is disclosed herein. In anotheraspect, a method for decreasing echogenicity of a stretch mark isdisclosed herein. In some aspects, a method for inhibiting COX-1, MMP1,MMP3, MMP9, and/or tyrosinase activity is disclosed herein. In someaspects, a method for increasing collagen production is disclosedherein. In some aspects, a method for inhibiting oxidation of the skinand/or providing an antioxidant to the skin and/or a formulation isdisclosed herein. In some aspects, the methods include applying any oneof the topical compositions described herein to a stretch mark and/or toskin. In some aspects, the methods include applying any one of thetopical compositions described herein to a stretch mark and/or to skinat least twice a day. In some aspects, the methods include applying thecomposition to a stretch mark, a scar, a fine line, and/or a wrinkle.

In particular aspects, the compositions of the present invention areformulated as a topical skin composition. The composition can have adermatologically acceptable vehicle or carrier for the compounds andextracts. The composition can further include a moisturizing agent or ahumectant, a surfactant, a silicone containing compounds, a UV agent, anoil, and/or other ingredients identified in this specification or thoseknown in the art. The composition can be a lotion, cream, gel, serum,emulsion (e.g., oil-in-water, water-in-oil, silicone-in-water,water-in-silicone, water-in-oil-in-water, oil-in-water-in-oil,oil-in-water-in-silicone, etc.), solutions (e.g., aqueous orhydro-alcoholic solutions), anhydrous bases (e.g., lipstick or apowder), ointments, milk, paste, aerosol, solid forms, eye jellies, gelserums, gel emulsions, etc. The composition can be in powdered form(e.g., dried, lyophilized, particulate, etc.). The composition can beformulated for topical skin application at least 1, 2, 3, 4, 5, 6, 7, ormore times a day during use. In other aspects of the present invention,compositions can be storage stable or color stable, or both. It is alsocontemplated that the viscosity of the composition can be selected toachieve a desired result, e.g., depending on the type of compositiondesired, the viscosity of such composition can be from about 1 cps towell over 1 million cps or any range or integer derivable therein (e.g.,2 cps, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100,200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000,6000, 7000, 8000, 9000, 10000, 20000, 30000, 40000, 50000, 60000, 70000,80000, 90000, 100000, 200000, 300000, 400000, 500000, 600000, 700000,800000, 900000, 1000000, 2000000, 3000000, 4000000, 5000000, 10000000,cps, etc., as measured on a Brookfield Viscometer using a TC spindle at2.5 rpm at 25° C.).

The compositions in non-limiting aspects can have a pH of about 6 toabout 9. In other aspects, the pH can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, or 14. The compositions can include a triglyceride.Non-limiting examples include small, medium, and large chaintriglycerides. In certain aspects, the triglyceride is a medium chaintriglyceride (e.g., caprylic capric triglyceride). The compositions canalso include preservatives. Non-limiting examples of preservativesinclude methylparaben, propylparaben, or a mixture of methylparaben andpropylparaben. In some embodiments, the composition is paraben-free.

Compositions of the present invention can have UVA and UVB absorptionproperties. The compositions can have an sun protection factor (SPF) of2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45,50, 55, 60, or more, or any integer or derivative therein. Thecompositions can be sunscreen lotions, sprays, or creams.

The compositions of the present invention can also include any one of,any combination of, or all of the following additional ingredients:water, a chelating agent, a moisturizing agent, a preservative, athickening agent, a silicone containing compound, an essential oil, astructuring agent, a vitamin, a pharmaceutical ingredient, or anantioxidant, or any combination of such ingredients or mixtures of suchingredients. In certain aspects, the composition can include at leasttwo, three, four, five, six, seven, eight, nine, ten, or all of theseadditional ingredients identified in the previous sentence. Non-limitingexamples of these additional ingredients are identified throughout thisspecification and are incorporated into this section by reference. Theamounts of such ingredients can range from 0.0001% to 99.9% by weight orvolume of the composition, or any integer or range in between asdisclosed in other sections of this specification, which areincorporated into this paragraph by reference.

Kits that include the compositions of the present invention are alsocontemplated. In certain embodiments, the composition is comprised in acontainer. The container can be a bottle, dispenser, or package. Thecontainer can dispense a pre-determined amount of the composition. Incertain aspects, the compositions is dispensed in a spray, mist, dollop,or liquid. The container can include indicia on its surface. The indiciacan be a word, an abbreviation, a picture, or a symbol.

It is also contemplated that the compositions disclosed throughout thisspecification can be used as a leave-on or rinse-off composition. By wayof example, a leave-on composition can be one that is topically appliedto skin and remains on the skin for a period of time (e.g., at least 5,6, 7, 8, 9, 10, 20, or 30 minutes, or at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours,or overnight or throughout the day). Alternatively, a rinse-offcomposition can be a product that is intended to be applied to the skinand then removed or rinsed from the skin (e.g., with water) within aperiod of time such as less than 5, 4, 3, 2, or 1 minute. An example ofa rinse of composition can be a skin cleanser, shampoo, conditioner, orsoap. An example of a leave-on composition can be a skin moisturizer,sunscreen, mask, overnight cream, or a day cream.

It is contemplated that any embodiment discussed in this specificationcan be implemented with respect to any method or composition of theinvention, and vice versa. Furthermore, compositions of the inventioncan be used to achieve methods of the invention.

In one embodiment, compositions of the present invention can bepharmaceutically or cosmetically elegant or can have pleasant tactileproperties. “Pharmaceutically elegant,” “cosmetically elegant,” and/or“pleasant tactile properties” describes a composition that hasparticular tactile properties which feel pleasant on the skin (e.g.,compositions that are not too watery or greasy, compositions that have asilky texture, compositions that are non-tacky or sticky, etc.).Pharmaceutically or cosmetically elegant can also relate to thecreaminess or lubricity properties of the composition or to the moistureretaining properties of the composition.

Also contemplated is a product comprising a composition of the presentinvention. In non-limiting aspects, the product can be a cosmeticproduct. The cosmetic product can be those described in other sectionsof this specification or those known to a person of skill in the art.

Non-limiting examples of products include a moisturizer, a cream, alotion, a skin softener, a gel, a wash, a foundation, a night cream, alipstick, a cleanser, a toner, a sunscreen, a mask, an anti-agingproduct, a deodorant, an antiperspirant, a perfume, a cologne, etc.

Also disclosed are the following Embodiments 1 to 35 of the presentinvention. Embodiment 1 is a method of reducing the appearance of astretch mark in a person's skin, the method comprising topicallyapplying to the stretch mark an effective amount of a topicalcomposition comprising vegetable amino acids, Myrciaria dubia fruitextract, Peucedanum graveolens extract, and tetrahexyldecyl ascorbate,wherein the appearance of the stretch mark is reduced. Embodiment 2 isthe method of Embodiment 1, wherein the vegetable amino acids are anaqueous extract of navy bean, wherein the Myrciaria dubia fruit extractis an aqueous extract that comprises the pulp of Myrciaria dubia fruit,and wherein the Peucedanum graveolens extract is an aqueous extract.Embodiment 3 is the method of any of Embodiments 1 to 2, wherein theeffective amount of the topical composition increases collagen density,increases fibrillin-1 expression, decreases collagen networkorientation, inhibits COX-1, MMP1, MMP3, MMP9, and/or tyrosinaseactivity, increases collagen production, and/or provides an antioxidantin the skin having the stretch mark. Embodiment 4 is the method of anyof Embodiments 1 to 3, wherein the topical composition comprises 0.01 to5% w/w of vegetable amino acids, 0.01 to 5% w/w of Myrciaria dubia fruitextract, 0.01 to 3% w/w of Peucedanum graveolens extract, and 0.01 to 1%w/w of tetrahexyldecyl ascorbate. Embodiment 5 is the method of any ofEmbodiments 1 to 4, wherein the topical composition further compriseswater. Embodiment 6 is the method of Embodiment 5, wherein the topicalcomposition comprises 60 to 85% w/w of water. Embodiment 7 is the methodof any of Embodiments 1 to 6, wherein the topical composition furthercomprises pentylene glycol, cyclopentasiloxane, glycerin, andhydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer.Embodiment 8 is the method of Embodiment 7, wherein the topicalcomposition comprises 1 to 10% w/w of pentylene glycol, 1 to 5% w/w ofcyclopentasiloxane, 0.1 to 5% w/w of glycerin, and 0.1 to 5% w/w ofhydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer.Embodiment 9 is the method of any of Embodiments 1 to 8, wherein thetopical composition further comprises dimethicone, dimethiconol,phenoxyethanol, and ethylhexylglycerin. Embodiment 10 is the method ofany of Embodiments 1 to 9, wherein the topical composition furthercomprises PEG-12 dimethicone, triethanolamine, and disodium EDTA.Embodiment 11 is the method of Embodiment 10, wherein the topicalcomposition comprises 0.1 to 3% w/w of PEG-12 dimethicone, 0.1 to 3% w/wof triethanolamine, 0.01 to 1% w/w of disodium EDTA. Embodiment 12 isthe method of any of Embodiments 1 to 11, wherein the topicalcomposition is an emulsion, serum, gel, gel emulsion, or gel serum.Embodiment 13 is the method of any of Embodiments 1 to 12, wherein thetopical composition is applied to a stretch mark at least twice a day.Embodiment 14 is a topical composition comprising vegetable amino acids,Myrciaria dubia fruit extract, Peucedanum graveolens extract, andtetrahexyldecyl ascorbate. Embodiment 15 is the topical composition ofEmbodiment 14, wherein the vegetable amino acids are an aqueous extractof navy bean, wherein the Myrciaria dubia fruit extract is an aqueousextract that comprises the pulp of Myrciaria dubia fruit, and whereinthe Peucedanum graveolens extract is an aqueous extract. Embodiment 16is the topical composition of any of Embodiments 14 to 15, wherein thetopical composition is capable of increasing collagen density,increasing fibrillin-1 expression, and/or decreases collagen networkorientation in skin having a stretch mark. Embodiment 17 is the topicalcomposition of any of Embodiments 14 to 16, wherein the topicalcomposition is capable of inhibiting COX-1, MMP1, MMP3, MMP9, and/ortyrosinase activity, increasing collagen production, and/or providing anantioxidant in skin. Embodiment 18 is the topical composition ofEmbodiment 17, wherein the topical composition comprises an effectiveamount of Myrciaria dubia fruit extract to inhibit COX-1, MMP1, MMP3,MMP9, and/or tyrosinase activity, increase collagen production, and/orprovide an antioxidant in skin. Embodiment 19 is the topical compositionof any of Embodiments 14 to 18, comprising 0.1 to 5% w/w of vegetableamino acids, 0.1 to 5% w/w of Myrciaria dubia fruit extract, 0.1 to 3%w/w of Peucedanum graveolens extract, and 0.01 to 1% w/w oftetrahexyldecyl ascorbate. Embodiment 20 is the topical composition ofany of Embodiments 14 to 19, further comprising water. Embodiment 21 isthe topical composition of Embodiment 20, comprising 60 to 85% w/w ofwater. Embodiment 22 is the topical composition of any of Embodiments 14to 21, further comprising pentylene glycol, cyclopentasiloxane,glycerin, and hydroxyethyl acrylate/sodium acryloyldimethyl tauratecopolymer. Embodiment 23 is the topical composition of Embodiment 22,comprising 1 to 10% w/w of pentylene glycol, 1 to 5% w/w ofcyclopentasiloxane, 0.1 to 5% w/w of glycerin, and 0.1 to 5% w/w ofhydroxyethyl acrylate/sodium acryloyldimethyl taurate copolymer.Embodiment 24 is the topical composition of any of Embodiments 14 to 23,further comprising dimethicone, dimethiconol, phenoxyethanol, andethylhexylglycerin. Embodiment 25 is the topical composition of any ofEmbodiments 14 to 24, further comprising PEG-12 dimethicone,triethanolamine, and disodium EDTA. Embodiment 26 is the topicalcomposition of Embodiment 25, comprising 0.1 to 3% w/w of PEG-12dimethicone, 0.1 to 3% w/w of triethanolamine, 0.01 to 1% w/w ofdisodium EDTA. Embodiment 27 is the topical composition of any ofEmbodiments 14 to 26, wherein the topical composition is an emulsion,serum, gel, gel emulsion, or gel serum. Embodiment 28 is a method ofimproving the condition of or treating skin comprising applying aneffective amount of the topical composition of any of Embodiments 14 to27 to skin. Embodiment 29 is the method of Embodiment 28, wherein theeffective amount of the topical composition increases collagen density,increases fibrillin-1 expression, and/or decreases collagen networkorientation in the skin. Embodiment 30 is the method of any ofEmbodiments 28 to 29, wherein the effective amount of the topicalcomposition decreases stretch mark visibility, improves the overallappearance of a stretch mark, decreases redness, increases smoothness,increases skin tone/evenness, increases elasticity, increases firmness,improves texture, decreases thickness of a stretch mark, and/ordecreases echogenicity of a stretch mark. Embodiment 31 is the method ofany of Embodiments 28 to 30, wherein the effective amount of the topicalcomposition inhibits COX-1, MMP1, MMP3, MMP9, and/or tyrosinaseactivity. Embodiment 32 is the method of any of Embodiments 28 to 31,wherein the effective amount of the topical composition increasescollagen production. Embodiment 33 is the method of any of Embodiments28 to 32, wherein the effective amount of the topical compositioninhibits oxidation of the skin and/or provides an antioxidant to theskin. Embodiment 34 is the method of any of Embodiments 28 to 33,wherein the topical composition is applied to skin at least twice a day.Embodiment 35 is the method of any one of Embodiments 28 to 34, whereinthe composition is applied to a stretch mark, a scar, a fine line,and/or a wrinkle.

“Topical application” means to apply or spread a composition onto thesurface of lips or keratinous tissue. “Topical skin composition”includes compositions suitable for topical application on skin and/orkeratinous tissue. Such compositions are typicallydermatologically-acceptable in that they do not have undue toxicity,incompatibility, instability, allergic response, and the like, whenapplied to skin and/or keratinous tissue. Topical skin care compositionsof the present invention can have a selected viscosity to avoidsignificant dripping or pooling after application to skin and/orkeratinous tissue.

“Keratinous tissue” includes keratin-containing layers disposed as theoutermost protective covering of mammals and includes, but is notlimited to, lips, skin, hair, and nails.

The term “about” or “approximately” are defined as being close to asunderstood by one of ordinary skill in the art. In one non-limitingembodiment the terms are defined to be within 10%, preferably within 5%,more preferably within 1%, and most preferably within 0.5%.

The term “substantially” and its variations are refers to ranges within10%, within 5%, within 1%, or within 0.5%.

The terms “inhibiting” or “reducing” or any variation of these termsincludes any measurable decrease or complete inhibition to achieve adesired result. The terms “promote” or “increase” or any variation ofthese terms includes any measurable increase or production of a proteinor molecule (e.g., matrix proteins such as collagen) to achieve adesired result.

The term “effective,” as that term is used in the specification and/orclaims, means adequate to accomplish a desired, expected, or intendedresult.

The use of the word “a” or “an” when used in conjunction with the terms“comprising,” “including,” “having,” or “containing,” or any variationsof these terms, in the claims and/or the specification may mean “one,”but it is also consistent with the meaning of “one or more,” “at leastone,” and “one or more than one.”

As used in this specification and claim(s), the words “comprising” (andany form of comprising, such as “comprise” and “comprises”), “having”(and any form of having, such as “have” and “has”), “including” (and anyform of including, such as “includes” and “include”) or “containing”(and any form of containing, such as “contains” and “contain”) areinclusive or open-ended and do not exclude additional, unrecitedelements or method steps.

The compositions and methods for their use can “comprise,” “consistessentially of,” or “consist of” any of the ingredients or stepsdisclosed throughout the specification. With respect to the phrase“consisting essentially of,” a basic and novel property of thecompositions and methods of the present invention is the ability toreduce the appearance of stretch marks on skin.

Other objects, features, and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and the examples,while indicating specific embodiments of the invention, are given by wayof illustration only. Additionally, it is contemplated that changes andmodifications within the spirit and scope of the invention will becomeapparent to those skilled in the art from this detailed description.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

As noted above, several of the unique aspects of the present inventionare to combine in a topical cosmetic composition vegetable amino acids,Myrciaria dubia fruit extract, Peucedanum graveolens extract, and/ortetrahexyldecyl ascorbate. This allows for the benefits of treating orreducing the appearance of striae on skin and/or preventing theformation of striae on skin. In some instances, the composition caninhibit COX-1, MMP1, MMP3, MMP9, and/or tyrosinase activity, can inducecollagen production, and/or can have an antioxidant capacity and/orprovide antioxidant benefits.

The following subsections describe non-limiting aspects of the presentinvention in further detail.

A particular composition of the present invention is designed to work asa topical composition. The composition relies on a unique combination ofany one of, any combination of, or all of vegetable amino acids,Myrciaria dubia fruit extract, Peucedanum graveolens extract, and/ortetrahexyldecyl ascorbate. A non-limiting examples of such a compositionis provided in Example 1, Table 1.

The compositions disclosed herein can be applied to the skin and remainon the skin for a period of time (e.g., at least 1, 2, 3, 4, 5, 10, 20,30, or 60 minutes or more). After which, the composition, if needed, canbe rinsed from the skin or peeled from the skin.

These and other non-limiting aspects of the present invention aredescribed in the following sections.

A. Active Ingredients

The present invention is premised on a determination that a combinationof active ingredients—vegetable amino acids, Myrciaria dubia fruitextract, Peucedanum graveolens extract, and/or tetrahexyldecylascorbate—can be used to improve the skin's visual appearance and reducethe appearance of, prevent, and/or treat striae. The combination ofingredients have been shown herein to decrease collagen networkorientation, increase collagen density, and increase fibrillin-1expression in stretch marks. Further, the combination was also shown toincrease collagen density and fibrillin-1 expression in skin that doesnot have stretch marks. In clinical studies, the combination was alsoshown to decrease visibility, increase overall appearance, decreaseredness, increase smoothness, increase skin tone/evenness, increaseelasticity, increase firmness, improve texture, decrease the thickness,and/or decrease echogenicity of people's stretch marks.

This combination of ingredients can be used in different products totreat various skin conditions. By way of non-limiting examples, thecombination of ingredients can be formulated in an emulsion, a gel, agel emulsion, a serum, a gel serum, a lotion, or a body butter.

Vegetable amino acids are biological organic compounds composed of amineand carboxylic acid functional groups, along with a side-chain specificto each amino acid. In some instances, vegetable amino acids are anaqueous extract of navy bean (Phaseolus vulgaris). Such an aqueousextract can be obtained from Carrubba Inc. under the trade name NavyBean. Therefore, an aqueous extract of navy beans can be an extract(powder or liquid in form) from navy beans that includes amino acidsfrom said navy beans. The extractant used can be water (aqueous extract)or a combination of water and alcohol (aqueous-alcoholic orhydro-alcoholic extract). In some instances, the vegetable amino acidsare an alcoholic extract of navy bean. The extractant used for analcoholic extract can be pure alcohol or a combination of alcohol andwater.

Myrciaria dubia fruit extract is an extract of the fruit pulp ofMyrciaria dubia, also known as camu camu. Myrciaria dubia is a smallbushy river side tree from the Amazon Rainforest vegetation in Peru andBrazil, which bears a red/purple cherry-like fruit. Camu camu fruit isrich in vitamin C. In some instances, Myrciaria dubia fruit extract isan aqueous extract that contains the pulp of Myrciaria dubia fruit.Myrciaria dubia fruit extract is commercially available and can beobtained from AMAX under the trade name Camu Camu or from Naturex underthe trade name Camu Camu LW. Therefore, an aqueous extract of Myrciariadubia fruit can be an extract (powder or liquid in form) from the pulpportion of this fruit. The extractant used can be water (aqueousextract) or a combination of water and alcohol (aqueous-alcoholic orhydro-alcoholic extract). In some instances, the Myrciaria dubia fruitextract is an alcoholic extract of Myrciaria dubia fruit. The extractantused for an alcoholic extract can be pure alcohol or a combination ofalcohol and water. In some instances, the Myrciaria dubia fruit extractcontains pulp or an extract of the pulp and alcohol, water, and ascorbicacid.

Peucedanum graveolens extract is the extract of Peucedanum graveolens,also known as Anethum graveolens or dill. Peucedanum graveolens is anannual herb. In some instances, Peucedanum graveolens extract is anaqueous extract. Peucedanum graveolens extract is commercially availableand can be obtained from BASF under the trade name LYS′LASTINE™.Therefore, an aqueous extract of Peucedanum graveolens can be an extract(powder or liquid in form) from a portion of or the whole plant ofPeucedanum graveolens. The extractant used can be water (aqueousextract) or a combination of water and alcohol (aqueous-alcoholic orhydro-alcoholic extract). In some instances, the Peucedanum graveolensextract is an alcoholic extract of Peucedanum graveolens. The extractantused for an alcoholic extract can be pure alcohol or a combination ofalcohol and water.

Tetrahexyldecyl ascorbate is also known as ascorbyl tetraisopalmitate.Tetrahexyldecyl ascorbate is a vitamin C derivative that functions as anantioxidant and skin conditioner agent. Tetrahexyldecyl ascorbate iscommercially available and can be obtained from Barnet under the tradename BV-OSC.

The extracts described herein can be extracts made through extractionmethods known in the art and combinations thereof. Non-limiting examplesof extraction methods include the use of liquid-liquid extraction, solidphase extraction, aqueous extraction, ethyl acetate, alcohol, acetone,oil, supercritical carbon dioxide, heat, pressure, pressure dropextraction, ultrasonic extraction, etc. Extracts can be a liquid, solid,dried liquid, re-suspended solid, etc.

B. Amounts of Ingredients

It is contemplated that the compositions of the present invention caninclude any amount of the ingredients discussed in this specification.The compositions can also include any number of combinations ofadditional ingredients described throughout this specification (e.g.,pigments, or additional cosmetic or pharmaceutical ingredients). Theconcentrations of the any ingredient within the compositions can vary.In non-limiting embodiments, for example, the compositions can comprise,consisting essentially of, or consist of, in their final form, forexample, at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%,0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%,0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%,0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%,0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%,0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%,0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0.0053%,0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%,0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%,0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%,0.0078%, 0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%,0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%,0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%,0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%,0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%,0.0650%, 0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%,0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%,0.1500%, 0.1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%,0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000%, 0.5250%,0.0550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%,0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%,0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%,1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%,3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%,4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%,5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%,6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%,7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%,9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 11%, 12%,13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%,27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 99% or any range derivable therein, of at least one of theingredients that are mentioned throughout the specification and claims.In non-limiting aspects, the percentage can be calculated by weight orvolume of the total composition. A person of ordinary skill in the artwould understand that the concentrations can vary depending on theaddition, substitution, and/or subtraction of ingredients in a givencomposition.

C. Vehicles

The compositions of the present invention can include or be incorporatedinto all types of vehicles and carriers. The vehicle or carrier can be apharmaceutically or dermatologically acceptable vehicle or carrier.Non-limiting examples of vehicles or carriers include water, glycerin,alcohol, oil, a silicon containing compound, a silicone compound, andwax. Variations and other appropriate vehicles will be apparent to theskilled artisan and are appropriate for use in the present invention. Incertain aspects, the concentrations and combinations of the compounds,ingredients, and agents can be selected in such a way that thecombinations are chemically compatible and do not form complexes whichprecipitate from the finished product.

D. Structure

The compositions of the present invention can be structured orformulated into a variety of different forms. Non-limiting examplesinclude emulsions (e.g., water-in-oil, water-in-oil-in-water,oil-in-water, silicone-in-water, water-in-silicone, oil-in-water-in-oil,oil-in-water-in-silicone emulsions), creams, lotions, solutions (bothaqueous and hydro-alcoholic), anhydrous bases (such as lipsticks andpowders), gels, masks, peels, and ointments. Variations and otherstructures will be apparent to the skilled artisan and are appropriatefor use in the present invention.

E. Additional Ingredients

In addition to the combination of ingredients disclosed by theinventors, the compositions can also include additional ingredients suchas cosmetic ingredients and pharmaceutical active ingredients.Non-limiting examples of these additional ingredients are described inthe following subsections.

1. Cosmetic Ingredients

The CTFA International Cosmetic Ingredient Dictionary and Handbook (2004and 2008) describes a wide variety of non-limiting cosmetic ingredientsthat can be used in the context of the present invention. Examples ofthese ingredient classes include: fragrance agents (artificial andnatural; e.g., gluconic acid, phenoxyethanol, and triethanolamine), dyesand color ingredients (e.g., Blue 1, Blue 1 Lake, Red 40, titaniumdioxide, D&C blue no. 4, D&C green no. 5, D&C orange no. 4, D&C red no.17, D&C red no. 33, D&C violet no. 2, D&C yellow no. 10, and D&C yellowno. 11), flavoring agents/aroma agents (e.g., Stevia rebaudiana(sweetleaf) extract, and menthol), adsorbents, lubricants, solvents,moisturizers (including, e.g., emollients, humectants, film formers,occlusive agents, and agents that affect the natural moisturizationmechanisms of the skin), water-repellants, UV absorbers (physical andchemical absorbers such as para-aminobenzoic acid (“PABA”) andcorresponding PABA derivatives, titanium dioxide, zinc oxide, etc.),essential oils, vitamins (e.g., A, B, C, D, E, and K), trace metals(e.g., zinc, calcium and selenium), anti-irritants (e.g., steroids andnon-steroidal anti-inflammatories), botanical extracts (e.g., Aloe vera,chamomile, cucumber extract, Ginkgo biloba, ginseng, and rosemary),anti-microbial agents, antioxidants (e.g., BHT and tocopherol),chelating agents (e.g., disodium EDTA and tetrasodium EDTA),preservatives (e.g., methylparaben and propylparaben), pH adjusters(e.g., sodium hydroxide and citric acid), absorbents (e.g., aluminumstarch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrin,talc, and zeolite), skin bleaching and lightening agents (e.g.,hydroquinone and niacinamide lactate), humectants (e.g., sorbitol, urea,methyl gluceth-20, saccharide isomerate, and mannitol), exfoliants,waterproofing agents (e.g., magnesium/aluminum hydroxide stearate), skinconditioning agents (e.g., aloe extracts, allantoin, bisabolol,ceramides, dimethicone, hyaluronic acid, biosaccharide gum-1,ethylhexylglycerin, pentylene glycol, hydrogenated polydecene,octyldodecyl oleate, and dipotassium glycyrrhizate). Non-limitingexamples of some of these ingredients are provided in the followingsubsections.

a. UV Absorption and/or Reflecting Agents

UV absorption and/or reflecting agents that can be used in combinationwith the compositions of the present invention include chemical andphysical sunblocks. Non-limiting examples of chemical sunblocks that canbe used include para-aminobenzoic acid (PABA), PABA esters (glycerylPABA, amyldimethyl PABA and octyldimethyl PABA), butyl PABA, ethyl PABA,ethyl dihydroxypropyl PABA, benzophenones (oxybenzone, sulisobenzone,benzophenone, and benzophenone-1 through 12), cinnamates (octylmethoxycinnamate, isoamyl p-methoxycinnamate, octylmethoxy cinnamate,cinoxate, diisopropyl methyl cinnamate, DEA-methoxycinnamate, ethyldiisopropylcinnamate, glyceryl octanoate dimethoxycinnamate and ethylmethoxycinnamate), cinnamate esters, salicylates (homomethyl salicylate,benzyl salicylate, glycol salicylate, isopropylbenzyl salicylate, etc.),anthranilates, ethyl urocanate, homosalate, octisalate, dibenzoylmethanederivatives (e.g., avobenzone), octocrylene, octyl triazone, digalloyltrioleate, glyceryl aminobenzoate, lawsone with dihydroxyacetone,ethylhexyl triazone, dioctyl butamido triazone, benzylidene malonatepolysiloxane, terephthalylidene dicamphor sulfonic acid, disodium phenyldibenzimidazole tetrasulfonate, diethylamino hydroxybenzoyl hexylbenzoate, bis diethylamino hydroxybenzoyl benzoate, bisbenzoxazoylphenyl ethylhexylimino triazine, drometrizole trisiloxane,methylene bis-benzotriazolyl tetramethylbutylphenol, andbis-ethylhexyloxyphenol methoxyphenyltriazine, 4-methylbenzylidenecamphor, and isopentyl 4-methoxycinnamate. Non-limiting examples ofphysical sunblocks include, kaolin, talc, petrolatum and metal oxides(e.g., titanium dioxide and zinc oxide).

b. Moisturizing Agents

Non-limiting examples of moisturizing agents that can be used with thecompositions of the present invention include amino acids, chondroitinsulfate, diglycerin, erythritol, fructose, glucose, glycerin, glycerolpolymers, glycol, 1,2,6-hexanetriol, honey, hyaluronic acid,hydrogenated honey, hydrogenated starch hydrolysate, inositol, lactitol,maltitol, maltose, mannitol, natural moisturizing factor, PEG-15butanediol, polyglyceryl sorbitol, salts of pyrrolidone carboxylic acid,potassium PCA, propylene glycol, saccharide isomerate, sodiumglucuronate, sodium PCA, sorbitol, sucrose, trehalose, urea, andxylitol.

Other examples include acetylated lanolin, acetylated lanolin alcohol,alanine, algae extract, Aloe barbadensis, Aloe barbadensis extract, Aloebarbadensis gel, Althea officinalis extract, apricot (Prunus armeniaca)kernel oil, arginine, arginine aspartate, Arnica montana extract,aspartic acid, avocado (Persea gratissima) oil, barrier sphingolipids,butyl alcohol, beeswax, behenyl alcohol, beta-sitosterol, birch (Betulaalba) bark extract, borage (Borago officinalis) extract, butcherbroom(Ruscus aculeatus) extract, butylene glycol, Calendula officinalisextract, Calendula officinalis oil, candelilla (Euphorbia cerifera) wax,canola oil, caprylic/capric triglyceride, cardamom (Elettariacardamomum) oil, carnauba (Copernicia cerifera) wax, carrot (Daucuscarota sativa) oil, castor (Ricinus communis) oil, ceramides, ceresin,ceteareth-5, ceteareth-12, ceteareth-20, cetearyl octanoate, ceteth-20,ceteth-24, cetyl acetate, cetyl octanoate, cetyl palmitate, chamomile(Anthemis nobilis) oil, cholesterol, cholesterol esters, cholesterylhydroxystearate, citric acid, clary (Salvia sclarea) oil, cocoa(Theobroma cacao) butter, coco-caprylate/caprate, coconut (Cocosnucifera) oil, collagen, collagen amino acids, corn (Zea mays) oil,fatty acids, decyl oleate, dimethicone copolyol, dimethiconol, dioctyladipate, dioctyl succinate, dipentaerythrityl hexacaprylate/hexacaprate,DNA, erythritol, ethoxydiglycol, ethyl linoleate, Eucalyptus globulusoil, evening primrose (Oenothera biennis) oil, fatty acids, Geraniummaculatum oil, glucosamine, glucose glutamate, glutamic acid,glycereth-26, glycerin, glycerol, glyceryl distearate, glycerylhydroxystearate, glyceryl laurate, glyceryl linoleate, glycerylmyristate, glyceryl oleate, glyceryl stearate, glyceryl stearate SE,glycine, glycol stearate, glycol stearate SE, glycosaminoglycans, grape(Vitis vinifera) seed oil, hazel (Corylus americana) nut oil, hazel(Corylus avellana) nut oil, hexylene glycol, hyaluronic acid, hybridsafflower (Carthamus tinctorius) oil, hydrogenated castor oil,hydrogenated coco-glycerides, hydrogenated coconut oil, hydrogenatedlanolin, hydrogenated lecithin, hydrogenated palm glyceride,hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenatedtallow glyceride, hydrogenated vegetable oil, hydrolyzed collagen,hydrolyzed elastin, hydrolyzed glycosaminoglycans, hydrolyzed keratin,hydrolyzed soy protein, hydroxylated lanolin, hydroxyproline, isocetylstearate, isocetyl stearoyl stearate, isodecyl oleate, isopropylisostearate, isopropyl lanolate, isopropyl myristate, isopropylpalmitate, isopropyl stearate, isostearamide DEA, isostearic acid,isostearyl lactate, isostearyl neopentanoate, jasmine (Jasminumofficinale) oil, jojoba (Buxus chinensis) oil, kelp, kukui (Aleuritesmoluccana) nut oil, lactamide MEA, laneth-16, laneth-10 acetate,lanolin, lanolin acid, lanolin alcohol, lanolin oil, lanolin wax,lavender (Lavandula angustifolia) oil, lecithin, lemon (Citrus medicalimonum) oil, linoleic acid, linolenic acid, Macadamia ternifolia nutoil, maltitol, matricaria (Chamomilla recutita) oil, methyl glucosesesquistearate, methylsilanol PCA, mineral oil, mink oil, mortierellaoil, myristyl lactate, myristyl myristate, myristyl propionate,neopentyl glycol dicaprylate/dicaprate, octyldodecanol, octyldodecylmyristate, octyldodecyl stearoyl stearate, octyl hydroxystearate, octylpalmitate, octyl salicylate, octyl stearate, oleic acid, olive (Oleaeuropaea) oil, orange (Citrus aurantium dulcis) oil, palm (Elaeisguineensis) oil, palmitic acid, pantethine, panthenol, panthenyl ethylether, paraffin, PCA, peach (Prunus persica) kernel oil, peanut (Arachishypogaea) oil, PEG-8 C12-18 ester, PEG-15 cocamine, PEG-150 distearate,PEG-60 glyceryl isostearate, PEG-5 glyceryl stearate, PEG-30 glycerylstearate, PEG-7 hydrogenated castor oil, PEG-40 hydrogenated castor oil,PEG-60 hydrogenated castor oil, PEG-20 methyl glucose sesquistearate,PEG-40 sorbitan peroleate, PEG-5 soy sterol, PEG-10 soy sterol, PEG-2stearate, PEG-8 stearate, PEG-20 stearate, PEG-32 stearate, PEG-40stearate, PEG-50 stearate, PEG-100 stearate, PEG-150 stearate,pentadecalactone, peppermint (Mentha piperita) oil, petrolatum,phospholipids, plankton extract, polyamino sugar condensate,polyglyceryl-3 diisostearate, polyquaternium-24, polysorbate 20,polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85,potassium myristate, potassium palmitate, propylene glycol, propyleneglycol dicaprylate/dicaprate, propylene glycol dioctanoate, propyleneglycol dipelargonate, propylene glycol laurate, propylene glycolstearate, propylene glycol stearate SE, PVP, pyridoxine dipalmitate,retinol, retinyl palmitate, rice (Oryza sativa) bran oil, RNA, rosemary(Rosmarinus officinalis) oil, rose oil, safflower (Carthamus tinctorius)oil, sage (Salvia officinalis) oil, sandalwood (Santalum album) oil,serine, serum protein, sesame (Sesamum indicum) oil, shea butter(Butyrospermum parkii), silk powder, sodium chondroitin sulfate, sodiumhyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodiumpolyglutamate, soluble collagen, sorbitan laurate, sorbitan oleate,sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol,soybean (Glycine soja) oil, sphingolipids, squalane, squalene,stearamide MEA-stearate, stearic acid, stearoxy dimethicone,stearoxytrimethylsilane, stearyl alcohol, stearyl glycyrrhetinate,stearyl heptanoate, stearyl stearate, sunflower (Helianthus annuus) seedoil, sweet almond (Prunus amygdalus dulcis) oil, synthetic beeswax,tocopherol, tocopheryl acetate, tocopheryl linoleate, tribehenin,tridecyl neopentanoate, tridecyl stearate, triethanolamine, tristearin,urea, vegetable oil, water, waxes, wheat (Triticum vulgare) germ oil,and ylang ylang (Cananga odorata) oil.

c. Antioxidants

Non-limiting examples of antioxidants that can be used with thecompositions of the present invention include acetyl cysteine, ascorbicacid polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanolpectinate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, cysteine, cysteine HCI, diamylhydroquinone,di-t-butylhydroquinone, dicetyl thiodipropionate, dioleyl tocopherylmethylsilanol, disodium ascorbyl sulfate, distearyl thiodipropionate,ditridecyl thiodipropionate, dodecyl gallate, erythorbic acid, esters ofascorbic acid, ethyl ferulate, ferulic acid, gallic acid esters,hydroquinone, isooctyl thioglycolate, kojic acid, magnesium ascorbate,magnesium ascorbyl phosphate, methylsilanol ascorbate, natural botanicalanti-oxidants such as green tea or grape seed extracts,nordihydroguaiaretic acid, octyl gallate, phenylthioglycolic acid,potassium ascorbyl tocopheryl phosphate, potassium sulfite, propylgallate, quinones, rosmarinic acid, sodium ascorbate, sodium bisulfite,sodium erythorbate, sodium metabisulfite, sodium sulfite, superoxidedismutase, sodium thioglycolate, sorbityl furfural, thiodiglycol,thiodiglycolamide, thiodiglycolic acid, thioglycolic acid, thiolacticacid, thiosalicylic acid, tocophereth-5, tocophereth-10, tocophereth-12,tocophereth-18, tocophereth-50, tocopherol, tocophersolan, tocopherylacetate, tocopheryl linoleate, tocopheryl nicotinate, tocopherylsuccinate, and tris(nonylphenyl)phosphite.

d. Structuring Agents

In other non-limiting aspects, the compositions of the present inventioncan include a structuring agent. Structuring agent, in certain aspects,assist in providing rheological characteristics to the composition tocontribute to the composition's stability. In other aspects, structuringagents can also function as an emulsifier or surfactant. Non-limitingexamples of structuring agents include stearic acid, palmitic acid,stearyl alcohol, cetyl alcohol, behenyl alcohol, stearic acid, palmiticacid, the polyethylene glycol ether of stearyl alcohol having an averageof about 1 to about 21 ethylene oxide units, the polyethylene glycolether of cetyl alcohol having an average of about 1 to about 5 ethyleneoxide units, and mixtures thereof.

e. Emulsifiers

In certain aspects of the present invention, the compositions do notinclude an emulsifier. In other aspects, however, the compositions caninclude one or more emulsifiers. Emulsifiers can reduce the interfacialtension between phases and improve the formulation and stability of anemulsion. The emulsifiers can be nonionic, cationic, anionic, andzwitterionic emulsifiers (See McCutcheon's (1986); U.S. Pat. Nos.5,011,681; 4,421,769; 3,755,560). Non-limiting examples include estersof glycerin, esters of propylene glycol, fatty acid esters ofpolyethylene glycol, fatty acid esters of polypropylene glycol, estersof sorbitol, esters of sorbitan anhydrides, carboxylic acid copolymers,esters and ethers of glucose, ethoxylated ethers, ethoxylated alcohols,alkyl phosphates, polyoxyethylene fatty ether phosphates, fatty acidamides, acyl lactylates, soaps, TEA stearate, DEA oleth-3 phosphate,polyethylene glycol 20 sorbitan monolaurate (polysorbate 20),polyethylene glycol 5 soya sterol, steareth-2, steareth-20, steareth-21,ceteareth-20, cetearyl glucoside, cetearyl alcohol, C12-13 pareth-3,PPG-2 methyl glucose ether distearate, PPG-5-ceteth-20,bis-PEG/PPG-20/20 dimethicone, ceteth-10, polysorbate 80, cetylphosphate, potassium cetyl phosphate, diethanolamine cetyl phosphate,polysorbate 60, glyceryl stearate, PEG-100 stearate, arachidyl alcohol,arachidyl glucoside, and mixtures thereof.

f. Silicone Containing Compounds

In non-limiting aspects, silicone containing compounds include anymember of a family of polymeric products whose molecular backbone ismade up of alternating silicon and oxygen atoms with side groupsattached to the silicon atoms. By varying the —Si—O— chain lengths, sidegroups, and crosslinking, silicones can be synthesized into a widevariety of materials. They can vary in consistency from liquid to gel tosolids.

The silicone containing compounds that can be used in the context of thepresent invention include those described in this specification or thoseknown to a person of ordinary skill in the art. Non-limiting examplesinclude silicone oils (e.g., volatile and non-volatile oils), gels, andsolids. In certain aspects, the silicon containing compounds includes asilicone oils such as a polyorganosiloxane. Non-limiting examples ofpolyorganosiloxanes include dimethicone, cyclomethicone,polysilicone-11, phenyl trimethicone, trimethylsilylamodimethicone,stearoxytrimethylsilane, or mixtures of these and other organosiloxanematerials in any given ratio in order to achieve the desired consistencyand application characteristics depending upon the intended application(e.g., to a particular area such as the skin, hair, or eyes). A“volatile silicone oil” includes a silicone oil have a low heat ofvaporization, i.e. normally less than about 50 cal per gram of siliconeoil. Non-limiting examples of volatile silicone oils include:cyclomethicones such as Dow Corning 344 Fluid, Dow Corning 345 Fluid,Dow Corning 244 Fluid, and Dow Corning 245 Fluid, Volatile Silicon 7207(Union Carbide Corp., Danbury, Conn.); low viscosity dimethicones, i.e.dimethicones having a viscosity of about 50 cst or less (e.g.,dimethicones such as Dow Corning 200-0.5 cst Fluid). The Dow CorningFluids are available from Dow Corning Corporation, Midland, Mich.Cyclomethicone and dimethicone are described in the Third Edition of theCTFA Cosmetic Ingredient Dictionary (incorporated by reference) ascyclic dimethyl polysiloxane compounds and a mixture of fully methylatedlinear siloxane polymers end-blocked with trimethylsiloxy units,respectively. Other non-limiting volatile silicone oils that can be usedin the context of the present invention include those available fromGeneral Electric Co., Silicone Products Div., Waterford, N.Y. and SWSSilicones Div. of Stauffer Chemical Co., Adrian, Mich.

g. Exfoliating Agent

Exfoliating agents include ingredients that remove dead skin cells onthe skin's outer surface. These agents may act through mechanical,chemical, and/or other means. Non-limiting examples of mechanicalexfoliating agents include abrasives such as pumice, silica, cloth,paper, shells, beads, solid crystals, solid polymers, etc. Non-limitingexamples of chemical exfoliating agents include acids and enzymeexfoliants. Acids that can be used as exfoliating agents include, butare not limited to, glycolic acid, lactic acid, citric acid, alphahydroxy acids, beta hydroxy acids, etc. Other exfoliating agents knownto those of skill in the art are also contemplated as being usefulwithin the context of the present invention.

h. Essential Oils

Essential oils include oils derived from herbs, flowers, trees, andother plants. Such oils are typically present as tiny droplets betweenthe plant's cells, and can be extracted by several method known to thoseof skill in the art (e.g., steam distilled, enfleurage (i.e., extractionby using fat), maceration, solvent extraction, or mechanical pressing).When these types of oils are exposed to air they tend to evaporate(i.e., a volatile oil). As a result, many essential oils are colorless,but with age they can oxidize and become darker. Essential oils areinsoluble in water and are soluble in alcohol, ether, fixed oils(vegetal), and other organic solvents. Typical physical characteristicsfound in essential oils include boiling points that vary from about 160°to 240° C. and densities ranging from about 0.759 to about 1.096.

Essential oils typically are named by the plant from which the oil isfound. For example, rose oil or peppermint oil are derived from rose orpeppermint plants, respectively. Non-limiting examples of essential oilsthat can be used in the context of the present invention include sesameoil, macadamia nut oil, tea tree oil, evening primrose oil, Spanish sageoil, Spanish rosemary oil, coriander oil, thyme oil, pimento berriesoil, rose oil, anise oil, balsam oil, bergamot oil, rosewood oil, cedaroil, chamomile oil, sage oil, clary sage oil, clove oil, cypress oil,eucalyptus oil, fennel oil, sea fennel oil, frankincense oil, geraniumoil, ginger oil, grapefruit oil, jasmine oil, juniper oil, lavender oil,lemon oil, lemongrass oil, lime oil, mandarin oil, marjoram oil, myrrhoil, neroli oil, orange oil, patchouli oil, pepper oil, black pepperoil, petitgrain oil, pine oil, rose otto oil, rosemary oil, sandalwoodoil, spearmint oil, spikenard oil, vetiver oil, wintergreen oil, orylang ylang. Other essential oils known to those of skill in the art arealso contemplated as being useful within the context of the presentinvention.

i. Thickening Agents

Thickening agents, including thickener or gelling agents, includesubstances which that can increase the viscosity of a composition.Thickeners includes those that can increase the viscosity of acomposition without substantially modifying the efficacy of the activeingredient within the composition. Thickeners can also increase thestability of the compositions of the present invention. In certainaspects of the present invention, thickeners include hydrogenatedpolyisobutene, trihydroxystearin, ammonium acryloyldimethyltaurate/vpcopolymer, or a mixture of them.

Non-limiting examples of additional thickening agents that can be usedin the context of the present invention include carboxylic acidpolymers, crosslinked polyacrylate polymers, polyacrylamide polymers,polysaccharides, and gums. Examples of carboxylic acid polymers includecrosslinked compounds containing one or more monomers derived fromacrylic acid, substituted acrylic acids, and salts and esters of theseacrylic acids and the substituted acrylic acids, wherein thecrosslinking agent contains two or more carbon-carbon double bonds andis derived from a polyhydric alcohol (see U.S. Pat. Nos. 5,087,445;4,509,949; 2,798,053; CTFA International Cosmetic Ingredient Dictionary,Fourth edition, 1991, pp. 12 and 80). Examples of commercially availablecarboxylic acid polymers include carbomers, which are homopolymers ofacrylic acid crosslinked with allyl ethers of sucrose or pentaerytritol(e.g., CARBOPOL™ 900 series from B. F. Goodrich).

Non-limiting examples of crosslinked polyacrylate polymers includecationic and nonionic polymers. Examples are described in U.S. Pat. Nos.5,100,660 ; 4,849,484; 4,835,206; 4,628,078; 4,599,379).

Non-limiting examples of polyacrylamide polymers (including nonionicpolyacrylamide polymers including substituted branched or unbranchedpolymers) include polyacrylamide, isoparaffin and laureth-7, multi-blockcopolymers of acrylamides and substituted acrylamides with acrylic acidsand substituted acrylic acids.

Non-limiting examples of polysaccharides include cellulose,carboxymethyl hydroxyethylcellulose, cellulose acetate propionatecarboxylate, hydroxyethylcellulose, hydroxyethyl ethylcellulose,hydroxypropylcellulose, hydroxypropyl methylcellulose, methylhydroxyethylcellulose, microcrystalline cellulose, sodium cellulosesulfate, and mixtures thereof. Another example is an alkyl substitutedcellulose where the hydroxy groups of the cellulose polymer ishydroxyalkylated (preferably hydroxy ethylated or hydroxypropylated) toform a hydroxyalkylated cellulose which is then further modified with aC10-C30 straight chain or branched chain alkyl group through an etherlinkage. Typically these polymers are ethers of C10-C30 straight orbranched chain alcohols with hydroxyalkylcelluloses. Other usefulpolysaccharides include scleroglucans comprising a linear chain of (1-3)linked glucose units with a (1-6) linked glucose every three unit.

Non-limiting examples of gums that can be used with the presentinvention include acacia, agar, algin, alginic acid, ammonium alginate,amylopectin, calcium alginate, calcium carrageenan, carnitine,carrageenan, dextrin, gelatin, gellan gum, guar gum, guarhydroxypropyltrimonium chloride, hectorite, hyaluronic acid, hydratedsilica, hydroxypropyl chitosan, hydroxypropyl guar, karaya gum, kelp,locust bean gum, natto gum, potassium alginate, potassium carrageenan,propylene glycol alginate, sclerotium gum, sodium carboxymethyl dextran,sodium carrageenan, tragacanth gum, xanthan gum, and mixtures thereof.

Preservatives

Non-limiting examples of preservatives that can be used in the contextof the present invention include quaternary ammonium preservatives suchas polyquaternium-1 and benzalkonium halides (e.g., benzalkoniumchloride (“BAC”) and benzalkonium bromide), parabens (e.g.,methylparabens and propylparabens), phenoxyethanol, benzyl alcohol,chlorobutanol, phenol, sorbic acid, thimerosal or combinations thereof.

2. Pharmaceutical Ingredients

Pharmaceutical active agents are also contemplated as being useful withthe compositions of the present invention. Non-limiting examples ofpharmaceutical active agents include anti-acne agents, agents used totreat rosacea, analgesics, anesthetics, anorectals, antihistamines,anti-inflammatory agents including non-steroidal anti-inflammatorydrugs, antibiotics, antifungals, antivirals, antimicrobials, anti-canceractives, scabicides, pediculicides, antineoplastics, antiperspirants,antipruritics, antipsoriatic agents, anti seborrheic agents,biologically active proteins and peptides, burn treatment agents,cauterizing agents, depigmenting agents, depilatories, diaper rashtreatment agents, enzymes, hair growth stimulants, hair growthretardants including DFMO and its salts and analogs, hemostatics,kerotolytics, canker sore treatment agents, cold sore treatment agents,dental and periodontal treatment agents, photosensitizing actives, skinprotectant/barrier agents, steroids including hormones andcorticosteroids, sunburn treatment agents, sunscreens, transdermalactives, nasal actives, vaginal actives, wart treatment agents, woundtreatment agents, wound healing agents, etc.

F. Kits

Kits are also contemplated as being used in certain aspects of thepresent invention. For instance, compositions of the present inventioncan be included in a kit. A kit can include a container. Containers caninclude a bottle, a metal tube, a laminate tube, a plastic tube, adispenser, a pressurized container, a barrier container, a package, acompartment, a lipstick container, a compact container, cosmetic pansthat can hold cosmetic compositions, or other types of containers suchas injection or blow-molded plastic containers into which thedispersions or compositions or desired bottles, dispensers, or packagesare retained. The kit and/or container can include indicia on itssurface. The indicia, for example, can be a word, a phrase, anabbreviation, a picture, or a symbol.

The containers can dispense a pre-determined amount of the composition.In other embodiments, the container can be squeezed (e.g., metal,laminate, or plastic tube) to dispense a desired amount of thecomposition. The composition can be dispensed as a spray, an aerosol, aliquid, a fluid, or a semi-solid. The containers can have spray, pump,or squeeze mechanisms. A kit can also include instructions for employingthe kit components as well the use of any other compositions included inthe container. Instructions can include an explanation of how to apply,use, and maintain the compositions.

EXAMPLES

The following examples are included to demonstrate preferred embodimentsof the invention. It should be appreciated by those of skill in the artthat the techniques disclosed in the examples which follow representtechniques discovered by the inventor to function well in the practiceof the invention, and thus can be considered to constitute preferredmodes for its practice. However, those of skill in the art should, inlight of the present disclosure, appreciate that many changes can bemade in the specific embodiments which are disclosed and still obtain alike or similar result without departing from the spirit and scope ofthe invention.

All of the compositions and methods disclosed and claimed herein can bemade and executed without undue experimentation in light of the presentdisclosure. While the compositions and methods of this invention havebeen described in terms of preferred embodiments, it will be apparent tothose of skill in the art that variations may be applied to thecompositions and methods and in the steps or in the sequence of steps ofthe method described herein without departing from the concept, spirit,and scope of the invention. More specifically, it will be apparent thatcertain agents which are both chemically and physiologically related maybe substituted for the agents described herein while the same or similarresults would be achieved. All such similar substitutes andmodifications apparent to those skilled in the art are deemed to bewithin the spirit, scope and concept of the invention as defined by theappended claims.

Example 1

Formulations having the ingredients from Example 1 were prepared astopical skin compositions. The formulation in Table 1 is an example of atopical skin composition prepared as a gel emulsion and/or a gel serum.

TABLE 1{circumflex over ( )} % Concentration Ingredient (by weight)Water 77 Pentylene Glycol 5 Cyclopentasiloxane 3 Glycerin 2 HydroxyethylAcrylate/Sodium 2 Acryloyldimethyl Taurate Copolymer DimethiconeSilicone HL 88* 2 DOW CORNING ® 1503** 2 Myrciaria dubia Fruit Extract 2Vegetable Amino Acids 2 PEG-12 Dimethicone 1 EUXYL ® PE 9010*** 1Peucedanum graveolens (Dill) Extract 1 Triethanolamine 0.4Tetrahexyldecyl Ascorbate 0.1 Disodium EDTA 0.1 Excipients{circumflexover ( )}{circumflex over ( )} q.s. {circumflex over ( )}Formulation canbe prepared by mixing the ingredients in a beaker under heat 70-75° C.until homogenous. Subsequently, the formulation can be cooled tostanding room temperature (20-25° C.). Further, and if desired,additional ingredients can be added, for example, to modify therheological properties of the composition. *Dimethicone Silicone HL 88contains dimethicone and is produced by Barnet Products Corp. **DOWCORNING ® 1503 is a blend of dimethicone and dimethiconol and isproduced by DOW CORNING ® ***EUXYL ® PE 9010 contains phenoxyethanol andethylhexylglycerin and is produced by Schülke Inc. {circumflex over( )}{circumflex over ( )}Excipients can be added, for example, to modifythe rheological properties of the composition. Alternatively, the amountof water can be varied so long as the amount of water in the compositionis at least 60% w/w, and preferably between 60 to 85% w/w.

Example 2 Activity on Stretch Marks on Human Living Skin Explants

It has been determined that the combination of vegetable amino acids,Myrciaria dubia fruit extract, Peucedanum graveolens extract, andtetrahexyldecyl ascorbate can decrease collagen network orientation,increase collagen density, and increase fibrillin-1 expression instretch marks. Further, it was determined that the combination increasescollagen density and fibrillin-1 expression in normal skin. The resultsdemonstrate that the combination increases the skin's repair activity ofstretch marks and skin. The assays used to test these properties of thecombination tested the formulation described above in Table 1 (“testcombination”) and utilized living human skin explants with stretchmarks.

Briefly, the assay tested the general morphology of human living skinexplants with and without stretch marks after Masson's trichromestaining and also tested fibrillin-1 expression by fibrillin-1immunostaining of the explants.

Human living skin explants—The skin explants were obtained from anabdoplasty from a 41-year old woman. 18 rectangular, skin explants(1×1.5 cm) with stretch marks and without (“normal”) were prepared andkept in BEM culture medium at 37° C. in a humid, 5% CO₂ atmosphere.

Treatment assay—For the test samples receiving test combination, on day0, 3, 5, and 7 two mg/cm² of the test combination was topically appliedon the surface of the explant samples by spreading with a spatula.Control explants did not receive any test combination or any othertreatment. Culture medium was refreshed on day 3, 5, and 7 for eachexplant. Each treatment and control was performed in triplicate. Toserve as a base line control, on day 0, three explants each of normalskin receiving no test composition and stretch mark skin receiving notest composition were collected and cut into two parts. One part wasfixed in buffered formalin and the other part was frozen at −80° C. Onday 10, three explants from each of: non-treated normal skin explant,non-treated stretch mark skin explant, test composition treated normalskin explant, and test composition treated stretch mark skin explantwere collected and cut into two parts. One part was fixed in bufferedformalin and the other part was frozen at −80° C.

Histological processing—The formalin fixed samples were fixed for 24hours in buffered formalin. Afterwards, the samples were dehydratedusing a Leica TP 1010 dehydration automat and impregnated in paraffinusing a Leica EG 1160 embedding station. The impregnated samples inparaffin were sectioned in 5 μ m thick sections using a Leica RM 2125Minot-type microtome. The sections were mounted on SUPERFROST®histological glass slides. The paraffinized sections were stainedaccording to Masson's trichrome, Goldner variant, for observation ofgeneral morphology. Morphology was observed under a microscope andrecorded by digital photography.

Frozen samples were cut into 7 μ m thick sections using a Leica CM 3050cryostat. The samples were then mounted on SUPERFROST® plus silanizedglass slides. The frozen sections were immunostained for the presence offibrillin-1 using a monoclonal anti-fibrillin-1 antibody, clone 11C.3(Novus Biological, NB110-8146) diluted at 1:500 in PBS-BSA 0.3% Tween 200.05%. Staining was performed for one hour at room temperature andenhanced with a streptavidin/biotin system and revealed using FITC(Invitrogen, SA 1001). Nuclei were post stained with propidium iodide.Staining was assessed by observation under a microscope and recorded bydigital photography.

Results—On day 10, the test combination showed restructuring activity onstretch marks by inducing an increase of collagen density in thepapillary dermis with stretch marks and a decrease of collagen networkorientation in comparison to controls. The test combination also inducedan increase of fibrillin-1 expression in the papillary dermis of theexplants with and without stretch marks in comparison with controls.

Example 3 Clinical Study

In clinical studies disclosed herein, it has been determined that thecombination of vegetable amino acids, Myrciaria dubia fruit extract,Peucedanum graveolens extract, and tetrahexyldecyl ascorbate candecrease visibility, beneficially increase overall appearance, decreaseredness, increase smoothness, increase skin tone/evenness, increaseelasticity, increase firmness, improve texture, decrease the thickness,and decrease echogenicity of subjects' stretch marks.

Briefly, 26 total subjects were divided into three groups. The subjectsranged in age from 23 to 45 years and all had hip, thigh, and/orabdominal stretch marks for between 0 and 36 months. Each subject wasprovided with the topical composition described in Table 1 abovecontaining a combination of vegetable amino acids, Myrciaria dubia fruitextract, Peucedanum graveolens extract, and tetrahexyldecyl ascorbate.The topical skin composition in Table 1 is designed to be a vehicle forthese active ingredients. This set-up allowed for confirmation that thecombination of these active ingredients (i.e., vegetable amino acids,Myrciaria dubia fruit extract, Peucedanum graveolens extract, andtetrahexyldecyl ascorbate) can reduce or decrease the appearance ofstretch marks on a person's skin. Reducing/decreasing the appearance ofstretch marks can be measured in the manner provided below.

The subjects were instructed to apply the composition to their stretchmarks twice daily for the duration of the 12 week study.

Evaluation—Subjects' stretch marks were evaluated at the baseline timeperiod and at week 3, 6, and 12 of treatment. An expert clinical graderevaluated the redness, texture/smoothness (visual and tactile), skintone/evenness, elasticity, visibility, and overall appearance of eachsubject's stretch marks at each time point. Firmness and elasticity werealso evaluated by a CUTOMETER® instrument, texture was also evaluated byuse of a VIVOSIGHT® instrument and also by use of Clarity Lite imaging,and echogenicity was evaluated using a DERMASCAN® instrument. Further,consumer perception was evaluated at week 3, 6, and 12 of treatment by asubjective questionnaire providing a 5 point scale for response to eachquestion, the 5 points on the scale were “Strongly Disagree,”“Disagree,” “Neutral,” “Agree,” and “Strongly Agree.” See Table 5.

Results—As shown in Tables 2 to 5 below expert clinical graderevaluation (Table 2), instrumental evaluation (Table 3 and Table 4), andconsumer perception (Table 5) all indicate that the compositioncontaining a combination of vegetable amino acids, Myrciaria dubia fruitextract, Peucedanum graveolens extract, and tetrahexyldecyl ascorbatestatistically significantly decreases visibility, beneficially increasesoverall appearance, decreases redness, increases smoothness, increasesskin tone/evenness, increases elasticity, increases firmness, improvestexture, decreases the thickness, and decreases echogenicity of stretchmarks. Some of these results were shown as early as at 3 weeks oftreatment with all showing statistical significance at week 12 andimprovement over the results of week 3 suggesting that even greaterbenefits from using the combination may be seen if used for more than 12weeks.

TABLE 2 Expert Clinical Grader Evaluation - Monadic, comparison toBaseline Anti-Stretch Mark Composition (Product A) Percent of MeanPercent Subjects Showing Time Point Improvement Improvement P-ValueAssessment (TX) n Mean ± SD From BL mean From BL TX vs. BL RednessBaseline (BL) 26 1.50 ± 0.58 Week 3 26 1.23 ± 0.81 21.15% 23.1% 0.016*Week 6 24 1.04 ± 0.80 32.63% 45.8% 0.002* Week 12 26 0.84 ± 0.78 47.43%57.7% <0.001* Texture/ Baseline 26 4.76 ± 1.00 Smoothness Week 3 26 3.50± 0.54 24.17% 84.6% <0.001* (Visual) Week 6 24 3.09 ± 0.95 29.37% 91.7%<0.001* Week 12 26 2.85 ± 0.69 37.54% 92.3% <0.001* Texture/ Baseline 264.60 ± 0.89 Smoothness Week 3 26 3.26 ± 0.54 26.46% 88.5% <0.001*(Tactile) Week 6 24 3.12 ± 0.88 28.31% 91.7% <0.001* Week 12 26 2.80 ±0.65 36.45% 92.3% <0.001* Skin Baseline 26 4.75 ± 0.83 Tone/EvennessWeek 3 26 3.58 ± 0.60 23.35% 96.2% <0.001* Week 6 24 3.15 ± 0.67 32.76% 100% <0.001* Week 12 26 2.77 ± 0.68 40.82% 96.2% <0.001* ElasticityBaseline 26 4.60 ± 0.75 (Tactile) Week 3 26 3.75 ± 0.55 17.29% 88.5%<0.001* Week 6 24 3.19 ± 0.67 29.36% 95.8% <0.001* Week 12 26 3.05 ±0.63 33.09%  100% <0.001* Stretch Mark Baseline 26 4.78 ± 0.92Visibility Week 3 26 3.74 ± 0.63 20.40% 88.5% <0.001* Week 6 24 3.42 ±1.02 25.68% 91.7% <0.001* Week 12 26 3.00 ± 0.67 36.51% 96.2% <0.001*Overall Baseline 26 4.91 ± 0.92 Appearance Week 3 26 3.73 ± 0.70 22.96%92.3% <0.001* Week 6 24 3.42 ± 0.79 28.00% 91.7% <0.001* Week 12 26 3.00± 0.77 38.33% 96.2% <0.001* *Indicates a statistically significantimprovement compared to baseline, p ≤ 0.05

TABLE 3 Instrumental Evaluation - Monadic, comparison to BaselineAnti-Stretch Mark Composition (Product A) Percent of Subjects Time MeanPercent Showing P-Value Point Improvement Improvement TX vs. Assessment(TX) n Mean ± SD From BL mean From BL BL Cutometer Firmness Baseline25{circumflex over ( )} 0.30 ± 0.06 R0 (Uf) (BL) Week 3 25{circumflexover ( )} 0.29 ± 0.06 0.19% 44.0% 0.521 Week 6 23{circumflex over ( )}0.26 ± 0.06 9.98% 65.2% 0.016* Week 12 25{circumflex over ( )} 0.26 ±0.06 10.08% 68.0% 0.011* Elasticity Baseline 25 0.79 ± 0.10 R5 (Ur/Ue)Week 3 25 0.89 ± 0.14 12.18% 80.0% 0.001* Week 6 23 0.88 ± 0.15 9.64%78.3% 0.005* Week 12 25 0.92 ± 0.14 16.22% 96.0% <0.001* VivoSightTexture (Ra) Baseline 26 0.01 ± 0.01 Week 3 25# 0.01 ± 0.00 NI 32.0%0.849 Week 6 24 0.01 ± 0.01 NI 37.5% 0.943 Week 12 26 0.02 ± 0.01 NI34.6% 0.096 Texture (Rz) Baseline 26 0.09 ± 0.03 Week 3 25# 0.17 ± 0.20101.36% 80.0% 0.049* Week 6 24 0.13 ± 0.07 62.33% 83.3% 0.037* Week 1226 0.22 ± 0.14 204.67% 61.5% <0.001* Dermascan Echo-genicity Baseline 25218.12 ± 42.67  Week 3 24 213.54 ± 40.65  3.00%  100% <0.001* Week 6 25198.00 ± 54.40  9.93%  100% 0.002* Week 12 25 188.52 ± 41.36  13.24% 100% <0.001* {circumflex over ( )}One subject (#36) did not haveCutometer data at baseline, and was not used for analysis (25 subject'sanalysis). #One subject (#36) did not have VivoSight data at Week 3 (25subjects analyzed). NI = No Improvement *Indicates a statisticallysignificant improvement compared to baseline, p ≤ 0.05 **Indicates astatistically significant worsening compared to baseline, p ≤ 0.05

TABLE 4 Clarity Lite Evaluation - Monadic, comparison to BaselineAnti-Stretch Mark Composition (Product A) Percent of Subjects Time MeanPercent Showing P-Value Point Improvement Improvement TX vs. Assessment(TX) n Mean ± SD From BL mean From BL BL Affected Stretch Mark TextureBaseline 23 57.78 ± 9.71 (BL) Week 3 23 61.39 ± 9.80 6.98% 52.2% 0.010*Week 6 21 63.19 ± 9.93 9.19% 76.2% 0.001* Week 12 23 68.04 ± 9.27 19.20%95.7% <0.001* Skin with No Stretch Marks Texture Baseline 23 61.22 ±9.91 Week 3 23 62.78 ± 7.24 3.96% 47.8% 0.162 Week 6 21 63.71 ± 4.526.40% 66.7% 0.174 Week 12 23 64.04 ± 5.68 6.49% 65.2% 0.109

TABLE 5 Subjective Questionnaire - Consumer Perception Week 3 Response n(%) Percent Strongly Strongly Responding Question n Agree Agree NeutralDisagree Disagree Favorably Anti-Stretch Mark Composition (Product A) 1.The test product improved 26 1 (3.8%) 16 6 3 0 (0.0%) 65.4%* thetexture/smoothness of (61.5%) (23.1%) (11.5%) stretch marks. 2. The testproduct improved 26 1 (3.8%) 9 10 6 0 (0.0%) 38.5% (decreased) thethickness of (34.6%) (38.5%) (23.1%) stretch marks. 3. The test productimproved 26 0 (0.0%) 16 8 2 0 (0.0%) 61.5%* the appearance of stretch(61.5%) (30.8%) (7.7%) marks. 4. The test product reduced the 26 0(0.0%) 11 13 2 0 (0.0%) 42.3% visibility of stretch marks. (42.3%)(50.0%) (7.7%) 5. The test product improved 26 0 (0.0%) 13 10 3 0 (0.0%)50.0% (lightened) the color of stretch (50.0%) (38.5%) (11.5%) marks. 6.The test product evened the 26 0 (0.0%) 11 12 3 0 (0.0%) 42.3% skin tone(color) of stretch (42.3%) (46.2%) (11.5%) marks. 7. The test productreduced the 26 1 (3.8%) 12 13 0 0 (0.0%) 50.0% redness of stretch marks.(46.2%) (50.0%) (0.0%) 8. The test product improved 26 1 (3.8%) 10 10 50 (0.0%) 42.3% the elasticity of skin. (38.5%) (38.5%) (19.2%) Week 6Response n (%) Percent Strongly Strongly Responding Question n AgreeAgree Neutral Disagree Disagree Favorably Anti-Stretch Mark Composition(Product A) 1. The test product improved 24 1 (4.2%) 19 4 0 (0.0%) 0(0.0%) 83.3%* the texture/smoothness of (79.2%) (16.7%) stretch marks.2. The test product improved 24 0 (0.0%) 18 5 1 (4.2%) 0 (0.0%) 75.0%*(decreased) the thickness of (75.0%) (20.8%) stretch marks. 3. The testproduct improved 24 2 (8.3%) 16 5 1 (4.2%) 0 (0.0%) 75.0%* theappearance of stretch (66.7%) (20.8%) marks. 4. The test product reducedthe 24 2 (8.3%) 15 7 0 (0.0%) 0 (0.0%) 70.8%* visibility of stretchmarks. (62.5%) (29.2%) 5. The test product improved 24 1 (4.2%) 16 7 0(0.0%) 0 (0.0%) 70.8%* (lightened) the color of stretch (66.7%) (29.2%)marks. 6. The test product evened the 24 1 (4.2%) 13 10 0 (0.0%) 0(0.0%) 58.3%* skin tone (color) of stretch (54.2%) (41.7%) marks. 7. Thetest product reduced the 24 1 (4.2%) 16 7 0 (0.0%) 0 (0.0%) 70.8%*redness of stretch marks. (66.7%) (29.2%) 8. The test product improved24 2 (8.3%) 14 8 0 (0.0%) 0 (0.0%) 66.7%* the elasticity of skin.(58.3%) (33.3%) Week 12 Response n (%) Percent Strongly StronglyResponding Question n Agree Agree Neutral Disagree Disagree FavorablyAnti-Stretch Mark Composition (Product A) 1. The test product improved26 6 16 4 0 (0.0%) 0 (0.0%) 84.6%* the texture/smoothness of (23.1%)(61.5%) (15.4%) stretch marks. 2. The test product improved 26 6 14 5 1(3.8%) 0 (0.0%) 76.9%* (decreased) the thickness of (23.1%) (53.8%)(19.2%) stretch marks. 3. The test product improved 26 5 15 4 2 (7.7%) 0(0.0%) 76.9%* the appearance of stretch (19.2%) (57.7%) (15.4%) marks.4. The test product reduced the 26 6 16 3 1 (3.8%) 0 (0.0%) 84.6%*visibility of stretch marks. (23.1%) (61.5%) (11.5%) 5. The test productimproved 26 6 17 3 0 (0.0%) 0 (0.0%) 88.5%* (lightened) the color ofstretch (23.1%) (65.4%) (11.5%) marks. 6. The test product evened the 266 15 5 0 (0.0%) 0 (0.0%) 80.8%* skin tone (color) of stretch (23.1%)(57.7%) (19.2%) marks. 7. The test product reduced the 26 7 15 3 1(3.8%) 0 (0.0%) 84.6%* redness of stretch marks. (26.9%) (57.7%) (11.5%)8. The test product improved 26 4 16 5 1 (3.8%) 0 (0.0%) 76.9%* theelasticity of skin. (15.4%) (61.5%) (19.2%) *Indicates the majority ofsubjects responded favorably, >50%

Example 4 In Vitro Assays

It has been determined that the Myrciaria dubia fruit extract (suppliedby Naturex under the trade name Camu Camu LW) can inhibit COX-1, MMP1,MMP3, MMP9, and tyrosinase activity, can induce collagen production, andhas an antioxidant capacity. A summary of results are found in Table 6and the methods used to determine the properties of the ingredients areprovided below.

TABLE 6 Assay Activity Collagen Production   +32% COX-1 Activity −90.92%MMP1 Activity −89.51% MMP3 Activity   −82% MMP9 Activity   −72%Tyrosinase Activity   −99% Antioxidant Capacity >0.3 mM Trolox

Collagen Stimulation Assay: Collagen is an extracellular matrix proteincritical for skin structure. Increased synthesis of collagen helpsimprove skin firmness and elasticity. This bioassay was used to examineeffects on the production of procollagen peptide (a precursor tocollagen) by human epidermal fibroblasts. The endpoint of this assay wasa spectrophotometric measurement that reflects the presence ofprocollagen peptide and cellular viability. The assay employed thequantitative sandwich enzyme immunoassay technique whereby a monoclonalantibody specific for procollagen peptide was pre-coated onto amicroplate. Standards and samples were pipetted into the wells and anyprocollagen peptide present was bound by the immobilized antibody. Afterwashing away any unbound substances, an enzyme-linked polyclonalantibody specific for procollagen peptide was added to the wells.Following a wash to remove any unbound antibody-enzyme reagent, asubstrate solution was added to the wells and color was developed inproportion to the amount of procollagen peptide bound in the initialstep. Color development was stopped and the intensity of the color at450 nm was measured using a microplate reader.

Cyclooxygenase 1 (COX-1) Assay: COX-1 contributes to the inflammatorypathway. COX is a bifunctional enzyme exhibiting both cyclooxygenase andperoxidase activities. The cyclooxygenase activity converts arachidonicacid to a hydroperoxy endoperoxide (Prostaglandin G2; PGG2) and theperoxidase component reduces the endoperoxide (Prostaglandin H2; PGH2)to the corresponding alcohol, the precursor of prostaglandins,thromboxanes, and prostacyclins. The peroxidase activity of COX-1 wasdetermined in the presence or absence of Myrciaria dubia fruit extractusing the Colorimetric COX (ovine) Inhibitor screening assay (#760111,Cayman Chemical).

COX-1 peroxidase activity was assayed colorimetrically by monitoring theappearance of oxidized N,N,N′,N′-tetramethyl-p-phenylenediamine (TMPD).This inhibitor screening assay includes both COX-1 and COX-2 enzymes inorder to screen isozyme-specific inhibitors. According to manufacturerinstructions, purified enzyme and heme with or without the test extractwere mixed in assay buffer and incubated with shaking for 15 min at roomtemperature. Following incubation, arachidonic acid and colorimetricsubstrate were added to initiate the reaction. Color progression wasevaluated by colorimetric plate reading at 590 nm. The percentinhibition of COX-1 activity was calculated and compared to non-treatedcontrols to determine the ability of test extracts to inhibit theactivity of the purified enzymes.

Inhibition of Matrix Metalloproteinase 1 Enzyme (MMP1): MMPs areextracellular proteases that play a role in many normal and diseasestates by virtue of their broad substrate specificity. MMP1 substratesinclude collagen IV. The activity of MMP1 in the presence or absenceofMyrciaria dubia fruit extract was determined using the MolecularProbes Enz/Chek Gelatinase/Collagenase Assay kit (#E12055).

Briefly, this kit utilizes a fluorogenic gelatin substrate to detectMMP1 protease activity in vitro. Upon proteolytic cleavage of thefluorogenic gelatin substrate, bright green fluorescence was revealedand was monitored using a fluorescent microplate reader to measureenzymatic activity. Test materials or control reagents were incubated inthe presence or absence of the purified enzyme and substrate todetermine their protease inhibitor capacity.

Matrix Metalloproteinase 3 and 9 Enzyme Activity (MMP3; MMP9) Assay:MMPs are extracellular proteases that play a role in many normal anddisease states by virtue of their broad substrate specificity. MMP3substrates include collagens, fibronectins, and laminin; while MMP9substrates include collagen VII, fibronectins and laminin. UsingColorimetric Drug Discovery kits from BioMol International for MMP3(AK-400) and MMP-9 (AK-410), this assay measured protease activity ofMMPs in the presence or absence of Myrciaria dubia fruit extract.Thiopeptide was used as a chromogenic substrate(Ac-PLG-[2-mercapto-4-methyl-pentanoyl]-LG-OC2H5)5,6. The MMP cleavagesite peptide bond is replaced by a thioester bond in the thiopeptide.Hydrolysis of this bond by an MMP produces a sulfhydryl group, whichreacts with DTNB [5,5′ -dithiobis(2-nitrobenzoic acid), Ellman'sreagent] to form 2-nitro-5-thiobenzoic acid, which was detected by itsabsorbance at 412 nm (ϵ=13,600 M-1 cm-1 at pH 6.0 and above 7).

Mushroom tyrosinase activity assay: In mammalian cells, tyrosinasecatalyzes two steps in the multi-step biosynthesis of melanin pigmentsfrom tyrosine (and from the polymerization of dopachrome). Tyrosinase islocalized in melanocytes and produces melanin (aromatic quinonecompounds) that imparts color to skin, hair, and eyes.

Tyrosinase activity was assayed by measuring the ability of purifiedmushroom tyrosinase (Sigma) to oxidize its substrate, L-Dopa (Fisher),in the presence or absence of Myrciaria dubia fruit extract. Oxidationof L-DOPA by the tyrosinase produced a pigment that was evaluated bycolorimetric plate reading at 490 nm. The percent inhibition of mushroomtyrosinase activity was calculated and compared to non-treated controlsto determine the ability of test ingredients to inhibit the activity ofpurified enzyme. Test inhibition was compared with that of the knowntyrosinase inhibitor kojic acid (Sigma).

For generation of samples and controls, subconfluent normal human adultepidermal fibroblasts (Cascade Biologics) were cultivated in standardDMEM growth medium with 10% fetal bovine serum (Mediatech) at 37° C. in10% CO2. The cells were treated with each of the tested ingredients andcontrols for 3 days. Following incubation, cell culture medium wascollected and the amount of Type I procollagen peptide secretion wasquantified using the sandwich enzyme linked immuno-sorbant assay (ELISA)from Takara (#MK101) as explained above.

Antioxidant Capacity: The antioxidant system of living organismsincludes enzymes such as superoxide dismutase, catalase, and glutathioneperoxidase; macromolecules such as albumin, ceruloplasmin, and ferritin;and an array of small molecules, including ascorbic acid, α-tocopherol,β-carotene, reduced glutathione, uric acid, and bilirubin. The sum ofendogenous and food-derived antioxidants represents the totalantioxidant activity of the extracellular fluid. Cooperation of all thedifferent antioxidants provides greater protection against attack byreactive oxygen or nitrogen radicals, than any single compound alone.Thus, the overall antioxidant capacity may give more relevant biologicalinformation compared to that obtained by the measurement of individualcomponents, as it accounts for the cumulative effect of all antioxidantspresent in plasma and body fluids.

Antioxidant capacity was determined by an Oxygen Radical Absorption (orAbsorbance) Capacity (ORAC) assay. This assay quantifies the degree andlength of time it takes to inhibit the action of an oxidizing agent,such as oxygen radicals, that are known to cause damage to cells (e.g.,skin cells). The ORAC value of the controls and Myrciaria dubia fruitextract was determined by the Zen-Bio ORAC Anti-oxidant Assay kit(#AOX-2). Briefly, this assay measures the loss of fluoresceinfluorescence over time due to the peroxyl-radical formation by thebreakdown of AAPH (2,2′-axobis-2-methyl propanimidamide,dihydrochloride). Trolox, a water soluble vitamin E analog, serves aspositive antioxidant control that inhibits fluorescein decay in a dosedependent manner.

Example 5 Additional Assays

Assays that can be used to determine the efficacy of any one of theingredients or any combination of ingredients or compositions havingsaid combination of ingredients disclosed throughout the specificationand claims can be determined by methods known to those of ordinary skillin the art. The following are non-limiting assays that can be used inthe context of the present invention. It should be recognized that othertesting procedures can be used, including, for example, objective andsubjective procedures.

B16 Pigmentation Assay: Melanogenesis is the process by whichmelanocytes produce melanin, a naturally produced pigment that impartscolor to skin, hair, and eyes. Inhibiting melanogenesis is beneficial toprevent skin darkening and lighten dark spots associated with aging.This bioassay utilizes B16-F1 melanocytes (ATCC), an immortalized mousemelanoma cell line, to analyze the effect of compounds on melanogenesis.The endpoint of this assay is a spectrophotometric measurement ofmelanin production and cellular viability. B16-F1 melanocytes, can becultivated in standard DMEM growth medium with 10% fetal bovine serum(Mediatech) at 37° C. in 10% CO₂ and then treated with any one of theactive ingredients, combination of ingredients, or compositions havingsaid combinations disclosed in the specification for 6 days. Followingincubation, melanin secretion is measured by absorbance at 405 nm andcellular viability is quantified.

Elastin Stimulation Assay: Elastin is a connective tissue protein thathelps skin resume shape after stretching or contracting. Elastin is alsoan important load-bearing protein used in places where mechanical energyis required to be stored. Elastin is made by linking many solubletropoelastin protein molecules, in a reaction catalyzed by lysyloxidase. Elastin secretion and elastin fibers can be monitored incultured human fibroblasts by staining of cultured human fibroblastsusing immunofluorescent antibodies directed against elastin.

Laminin and Fibronectin Stimulation Assay: Laminin and fibronectin aremajor proteins in the dermal-epidermal junction (DEJ) (also referred toas the basement membrane). The DEJ is located between the dermis and theepidermis interlocks forming fingerlike projections called rete ridges.The cells of the epidermis receive their nutrients from the bloodvessels in the dermis. The rete ridges increase the surface area of theepidermis that is exposed to these blood vessels and the needednutrients. The DEJ provides adhesion of the two tissue compartments andgoverns the structural integrity of the skin. Laminin and fibronectinare two structural glycoproteins located in the DEJ. Considered the gluethat holds the cells together, laminin and fibronectin are secreted bydermal fibroblasts to help facilitate intra- and inter-cellular adhesionof the epidermal calls to the DEJ. Laminin and fibronectin secretion canbe monitored by quantifying laminin and fibronectin in cell supernatantsof cultured human fibroblasts treated for 3 days with culture mediumwith or without 1.0% final concentration of the test ingredient(s).Following incubation, laminin and fibronectin content can be measuredusing immunofluorescent antibodies directed against each protein in anenzyme linked immuno-sorbant assay (ELISA). Measurements are normalizedfor cellular metabolic activity, as determined by bioconversion of3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS).

Tumor Necrosis Factor Alpha (TNF-α) Assay: The prototype ligand of theTNF superfamily, TNF-α, is a pleiotropic cytokine that plays a centralrole in inflammation. Increase in its expression is associated with anup regulation in pro-inflammatory activity. This bioassay can be used toanalyze the effect of any one of the active ingredients, combination ofingredients, or compositions having said combinations disclosed in thespecification on the production of TNF-α by human epidermalkeratinocytes. The endpoint of this assay can be a spectrophotometricmeasurement that reflects the presence of TNF-α and cellular viability.The assay employs the quantitative sandwich enzyme immunoassay techniquewhereby a monoclonal antibody specific for TNF-α has been pre-coatedonto a microplate. Standards and samples can be pipetted into the wellsand any TNF-α present is bound by the immobilized antibody. Afterwashing away any unbound substances, an enzyme-linked polyclonalantibody specific for TNF-α can be added to the wells. Following a washto remove any unbound antibody-enzyme reagent, a substrate solution canbe added to the wells and color develops in proportion to the amount ofTNF-α bound in the initial step using a microplate reader for detectionat 450 nm. The color development can be stopped and the intensity of thecolor can be measured. Subconfluent normal human adult keratinocytes(Cascade Biologics) cultivated in EpiLife standard growth medium(Cascade Biologics) at 37° C. in 5% CO₂, can be treated with phorbol12-myristate 13-acetate (PMA, 10 ng/ml, Sigma Chemical, #P1585-1MG) andany one of the active ingredients, combination of ingredients, orcompositions having said combinations disclosed in the specification for6 hours. PMA has been shown to cause a dramatic increase in TNF-αsecretion which peaks at 6 hours after treatment. Following incubation,cell culture medium can be collected and the amount of TNF-α secretionquantified using a sandwich enzyme linked immuno-sorbant assay (ELISA)from R&D Systems (#DTA00C).

Antioxidant (AO) Assay: An in vitro bioassay that measures the totalanti-oxidant capacity of any one of the ingredients, combination ofingredients, or compositions having said combinations disclosed in thespecification. The assay relies on the ability of antioxidants in thesample to inhibit the oxidation of ABTS®(2,2′-azino-di-[3-ethylbenzthiazoline sulphonate]) to ABTS®+bymetmyoglobin. The antioxidant system of living organisms includesenzymes such as superoxide dismutase, catalase, and glutathioneperoxidase; macromolecules such as albumin, ceruloplasmin, and ferritin;and an array of small molecules, including ascorbic acid, α-tocopherol,(3-carotene, reduced glutathione, uric acid, and bilirubin. The sum ofendogenous and food-derived antioxidants represents the totalantioxidant activity of the extracellular fluid. Cooperation of all thedifferent antioxidants provides greater protection against attack byreactive oxygen or nitrogen radicals, than any single compound alone.Thus, the overall antioxidant capacity may give more relevant biologicalinformation compared to that obtained by the measurement of individualcomponents, as it considers the cumulative effect of all antioxidantspresent in plasma and body fluids. The capacity of the antioxidants inthe sample to prevent ABTS oxidation is compared with that of Trolox, awater-soluble tocopherol analogue, and is quantified as molar Troloxequivalents. Anti-Oxidant capacity kit # 709001 from Cayman Chemical(Ann Arbor, Michigan USA) can be used as an in vitro bioassay to measurethe total anti-oxidant capacity of each of any one of the activeingredients, combination of ingredients, or compositions having saidcombinations disclosed in the specification. The protocol can befollowed according to manufacturer recommendations.

Lipoxygenase (LO) Assay: An in vitro lipoxygenase (LO) inhibition assay.LOs are non-heme iron-containing dioxygenases that catalyze the additionof molecular oxygen to fatty acids. Linoleate and arachidonate are themain substrates for LOs in plants and animals. Arachadonic acid may thenbe converted to hydroxyeicosotrienenoic (HETE) acid derivatives, thatare subsequently converted to leukotrienes, potent inflammatorymediators. This assay provides an accurate and convenient method forscreening lipoxygenase inhibitors by measuring the hydroperoxidesgenerated from the incubation of a lipoxygenase (5-, 12-, or 15-LO) witharachidonic acid. The Colorimetric LO Inhibitor screening kit (#760700,Cayman Chemical) can be used to determine the ability of each of theactive ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification toinhibit enzyme activity. Purified 15-lipoxygenase and test ingredientscan be mixed in assay buffer and incubated with shaking for 10 min atroom temperature. Following incubation, arachidonic acid can be added toinitiate the reaction and the mixtures can be incubated for anadditional 10 min at room temperature. Colorimetric substrate can beadded to terminate catalysis and color progression can be evaluated byfluorescence plate reading at 490 nm. The percent inhibition oflipoxyganse activity can be calculated compared to non-treated controlsto determine the ability of each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification to inhibit the activity of purifiedenzyme.

Elastase Assay: ENZCHEK® Elastase Assay (Kit# E-12056) from MolecularProbes (Eugene, Oreg. USA) can be used as an in vitro enzyme inhibitionassay for measuring inhibition of elastase activity for each of theactive ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification.The EnzChek kit contains soluble bovine neck ligament elastin that canbe labeled with dye such that the conjugate's fluorescence can bequenched. The non-fluorescent substrate can be digested by elastase orother proteases to yield highly fluorescent fragments. The resultingincrease in fluorescence can be monitored with a fluorescence microplatereader. Digestion products from the elastin substrate have absorptionmaxima at ˜505 nm and fluorescence emission maxima at ˜515 nm. Thepeptide, N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone, can beused as a selective, collective inhibitor of elastase when utilizing theEnzChek Elastase Assay Kit for screening for elastase inhibitors.

Oil Control Assay: An assay to measure reduction of sebum secretion fromsebaceous glands and/or reduction of sebum production from sebaceousglands can be assayed by using standard techniques known to those havingordinary skill in the art. In one instance, the forehead can be used.Each of the active ingredients, any one of the combination ofingredients, or compositions having said combinations disclosed in thespecification can be applied to one portion of the forehead once ortwice daily for a set period of days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, or more days), while another portion of the foreheadis not treated with the composition. After the set period of daysexpires, then sebum secretion can be assayed by application of fineblotting paper to the treated and untreated forehead skin. This is doneby first removing any sebum from the treated and untreated areas withmoist and dry cloths. Blotting paper can then be applied to the treatedand untreated areas of the forehead, and an elastic band can be placedaround the forehead to gently press the blotting paper onto the skin.After 2 hours the blotting papers can be removed, allowed to dry andthen transilluminated. Darker blotting paper correlates with more sebumsecretion (or lighter blotting paper correlates with reduced sebumsecretion.

Erythema Assay: An assay to measure the reduction of skin redness can beevaluated using a Minolta Chromometer. Skin erythema may be induced byapplying a 0.2% solution of sodium dodecyl sulfate on the forearm of asubject. The area is protected by an occlusive patch for 24 hrs. After24 hrs, the patch is removed and the irritation-induced redness can beassessed using the a* values of the Minolta Chroma Meter. The a* valuemeasures changes in skin color in the red region. Immediately afterreading, the area is treated with the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification. Repeat measurements can be taken atregular intervals to determine the formula's ability to reduce rednessand irritation.

Skin Moisture/Hydration Assay: Skin moisture/hydration benefits can bemeasured by using impedance measurements with the Nova Dermal PhaseMeter. The impedance meter measures changes in skin moisture content.The outer layer of the skin has distinct electrical properties. Whenskin is dry it conducts electricity very poorly. As it becomes morehydrated increasing conductivity results. Consequently, changes in skinimpedance (related to conductivity) can be used to assess changes inskin hydration. The unit can be calibrated according to instrumentinstructions for each testing day. A notation of temperature andrelative humidity can also be made. Subjects can be evaluated asfollows: prior to measurement they can equilibrate in a room withdefined humidity (e.g., 30-50%) and temperature (e.g., 68-72° C.). Threeseparate impedance readings can be taken on each side of the face,recorded, and averaged. The T5 setting can be used on the impedancemeter which averages the impedance values of every five secondsapplication to the face. Changes can be reported with statisticalvariance and significance. Each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification can be assayed according to this process.

Skin Clarity and Reduction in Freckles and Age Spots Assay: Skin clarityand the reduction in freckles and age spots can be evaluated using aMinolta Chromometer. Changes in skin color can be assessed to determineirritation potential due to product treatment using the a* values of theMinolta Chroma Meter. The a* value measures changes in skin color in thered region. This is used to determine whether each of the activeingredients, any one of the combination of ingredients, or compositionshaving said combinations disclosed in the specification is inducingirritation. The measurements can be made on each side of the face andaveraged, as left and right facial values. Skin clarity can also bemeasured using the Minolta Meter. The measurement is a combination ofthe a*, b, and L values of the Minolta Meter and is related to skinbrightness, and correlates well with skin smoothness and hydration. Skinreading is taken as above. In one non-limiting aspect, skin clarity canbe described as L/C where C is chroma and is defined as (a²+b²)^(1/2).

Skin Dryness, Surface Fine Lines, Skin Smoothness, and Skin Tone Assay:Skin dryness, surface fine lines, skin smoothness, and skin tone can beevaluated with clinical grading techniques. For example, clinicalgrading of skin dryness can be determined by a five point standardKligman Scale: (0) skin is soft and moist; (1) skin appears normal withno visible dryness; (2) skin feels slightly dry to the touch with novisible flaking; (3) skin feels dry, tough, and has a whitish appearancewith some scaling; and (4) skin feels very dry, rough, and has a whitishappearance with scaling. Evaluations can be made independently by twoclinicians and averaged.

Clinical Grading of Skin Tone Assay: Clinical grading of skin tone canbe performed via a ten point analog numerical scale: (10) even skin ofuniform, pinkish brown color. No dark, erythremic, or scaly patches uponexamination with a hand held magnifying lens. Microtexture of the skinvery uniform upon touch; (7) even skin tone observed withoutmagnification. No scaly areas, but slight discolorations either due topigmentation or erythema. No discolorations more than 1 cm in diameter;(4) both skin discoloration and uneven texture easily noticeable. Slightscaliness. Skin rough to the touch in some areas; and (1) uneven skincoloration and texture. Numerous areas of scaliness and discoloration,either hypopigmented, erythremic or dark spots. Large areas of unevencolor more than 1 cm in diameter. Evaluations were made independently bytwo clinicians and averaged.

Clinical Grading of Skin Smoothness Assay: Clinical grading of skinsmoothness can be analyzed via a ten point analog numerical scale: (10)smooth, skin is moist and glistening, no resistance upon dragging fingeracross surface; (7) somewhat smooth, slight resistance; (4) rough,visibly altered, friction upon rubbing; and (1) rough, flaky, unevensurface. Evaluations were made independently by two clinicians andaveraged.

Skin Smoothness and Wrinkle Reduction Assay With Methods Disclosed inPackman et al. (1978): Skin smoothness and wrinkle reduction can also beassessed visually by using the methods disclosed in Packman et al.(1978). For example, at each subject visit, the depth, shallowness andthe total number of superficial facial lines (SFLs) of each subject canbe carefully scored and recorded. A numerical score was obtained bymultiplying a number factor times a depth/width/length factor. Scoresare obtained for the eye area and mouth area (left and right sides) andadded together as the total wrinkle score.

Skin Firmness Assay with a Hargens Ballistometer: Skin firmness can bemeasured using a Hargens ballistometer, a device that evaluates theelasticity and firmness of the skin by dropping a small body onto theskin and recording its first two rebound peaks. The ballistometry is asmall lightweight probe with a relatively blunt tip (4 square mm-contactarea) was used. The probe penetrates slightly into the skin and resultsin measurements that are dependent upon the properties of the outerlayers of the skin, including the stratum corneum and outer epidermisand some of the dermal layers.

Skin Softness/Suppleness Assay with a Gas Bearing Electrodynamometer:Skin softness/suppleness can be evaluated using the Gas BearingElectrodynamometer, an instrument that measures the stress/strainproperties of the skin. The viscoelastic properties of skin correlatewith skin moisturization. Measurements can be obtained on thepredetermined site on the cheek area by attaching the probe to the skinsurface with double-stick tape. A force of approximately 3.5 gm can beapplied parallel to the skin surface and the skin displacement isaccurately measured. Skin suppleness can then be calculated and isexpressed as DSR (Dynamic Spring Rate in gm/mm).

Appearance of Lines and Wrinkles Assay with Replicas: The appearance oflines and wrinkles on the skin can be evaluated using replicas, which isthe impression of the skin's surface. Silicone rubber like material canbe used. The replica can be analyzed by image analysis. Changes in thevisibility of lines and wrinkles can be objectively quantified via thetaking of silicon replicas form the subjects' face and analyzing thereplicas image using a computer image analysis system. Replicas can betaken from the eye area and the neck area, and photographed with adigital camera using a low angle incidence lighting. The digital imagescan be analyzed with an image processing program and are of the replicascovered by wrinkles or fine lines was determined.

Surface Contour of the Skin Assay with a Profilometer/Stylus Method: Thesurface contour of the skin can be measured by using theprofilometer/Stylus method. This includes either shining a light ordragging a stylus across the replica surface. The vertical displacementof the stylus can be fed into a computer via a distance transducer, andafter scanning a fixed length of replica a cross-sectional analysis ofskin profile can be generated as a two-dimensional curve. This scan canbe repeated any number of times along a fix axis to generate a simulated3-D picture of the skin. Ten random sections of the replicas using thestylus technique can be obtained and combined to generate averagevalues. The values of interest include Ra which is the arithmetic meanof all roughness (height) values computed by integrating the profileheight relative to the mean profile height. Rt which is the maximumvertical distance between the highest peak and lowest trough, and Rzwhich is the mean peak amplitude minus the mean peak height. Values aregiven as a calibrated value in mm. Equipment should be standardizedprior to each use by scanning metal standards of know values. Ra Valuecan be computed by the following equation: R_(a)=Standardize roughness;l_(m)=the traverse (scan) length; and y=the absolute value of thelocation of the profile relative to the mean profile height (x-axis).

MELANODERM™ Assay: In other non-limiting aspects, the efficacy of eachof the active ingredients, any one of the combination of ingredients, orcompositions having said combinations disclosed in the specification canbe evaluated by using a skin analog, such as, for example, MELANODERM™.Melanocytes, one of the cells in the skin analog, stain positively whenexposed to L-dihydroxyphenyl alanine (L-DOPA), a precursor of melanin.The skin analog, MELANODERM™, can be treated with a variety of basescontaining each of the active ingredients, any one of the combination ofingredients, or compositions having said combinations disclosed in thespecification or with the base alone as a control. Alternatively, anuntreated sample of the skin analog can be used as a control.

Production of Filaggrin: Changes in the production of filaggrin inkeratinocytes due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Filaggrin is theprecursor to Natural Moisturizing Factor (NMF) in the skin. IncreasedNMF increases the moisture content of the skin. Filaggrin production intreated and non-treated keratinocytes can be determined using a bioassaythat analyzes filaggrin concentration in keratinocyte cell lysates. Anon-limiting example of a bioassay that can be used to quantifyfilaggrin production is the PROTEINSIMPLE® SIMON™ western blottingprotocol. For each sample, normal human epidermal keratinocytes (NHEK)are grown in EPI-200—Mattek EPILIFE® growth media with calcium from LifeTechnologies (M-EP-500-CA). NHEK are incubated in growth mediumovernight at 37° C. in 5% CO₂ prior to treatment. NHEK are thenincubated in growth medium with 1% test compound/extract or nocompound/extract (negative control) for 24 to 36 hours. The NHEK canthen be washed, collected, and stored on ice or colder until lysed onice using a lysis buffer and sonication. The protein concentrations ofthe samples can be determined and used to normalize the samples. Thelysates can be stored at −80° C. until use in the quantification assay.

The PROTEINSIMPLE® SIMON™ western blotting bioassay assay employs aquantitative western blotting immunoassay technique using an antibodyspecific for filaggrin to quantitatively detect filaggrin in the testsamples. Cell samples are lysed and normalized for proteinconcentration. Normalized samples and molecular weight standards canthen be loaded and ran on a denatured protein separation gel usingcapillary electrophoresis. The proteins in the gel are immobilized andimmunoprobed using a primary antibody specific for filaggrin. Theimmobilized proteins can then be immunoprobed with an enzyme-linkeddetection antibody that binds the primary antibody. A chemiluminescentsubstrate solution can then be added to the immobilized proteins toallow chemiluminescent development in proportion to the amount offilaggrin bound in the immobilization. The chemiluminescent developmentis stopped at a specific time and the intensity of the chemiluminescentsignal can be measured and compared to positive and negative controls.

Production of Occludin: Changes in the production of occludin inkeratinocytes due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Occludin is a proteincritical to the formulation of tight junctions and the skin's moisturebarrier function. A non-limiting example of how occludin production intreated and non-treated keratinocytes can be determined is by the use ofa bioassay that analyzes occludin concentration in keratinocyte celllysates. The bioassay can be performed using PROTEINSIMPLE® SIMON™western blotting protocol. For the samples, adult human epidermalkeratinocytes (HEKa) from Life Technologies (C-005-5C) can be grown at37° C. and 5% CO2 for 24 hours in Epilife growth media with calcium fromLife Technologies (M-EP-500-CA) supplemented with Keratinocyte GrowthSupplement (HKGS) from Life Technologies (S-101-5). HEKa are thenincubated in growth medium with test compound/extract, nocompound/extract for negative control, or with 1 mM CaCl₂ for positivecontrol for 24 to 48 hours. The HEKa are then washed, collected, andstored on ice or colder until lysed on ice using a lysis buffer andsonication. The protein concentrations of the samples can be determinedand used to normalize the samples. The lysates are stored at −80° C.until use in the bioassay.

The PROTEINSIMPLE® SIMON™ western blotting bioassay assay employs aquantitative western blotting immunoassay technique using an antibodyspecific for occludin to quantitatively detect occludin in the testsamples. Cell samples are lysed and normalized for proteinconcentration. Normalized samples and molecular weight standards arethen loaded and ran on a denatured protein separation gel usingcapillary electrophoresis. The proteins in the gel are then immobilizedand immunoprobed using a primary antibody specific for occludin. Theimmobilized proteins are immunoprobed with an enzyme-linked detectionantibody that binds the primary antibody. A chemiluminescent substratesolution is then added to the immobilized proteins to allowchemiluminescent development in proportion to the amount of occludinbound in the immobilization. The chemiluminescent development can bestopped at a specific time and the intensity of the chemiluminescentsignal can be measured and compared to positive and negative controls.

Keratinocyte Monolayer Permeability: Changes in the permeability of akeratinocyte monolayer due to each of the active ingredients, any one ofthe combination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Keratinocyte monolayerpermeability is a measure of skin barrier integrity. Keratinocytemonolayer permeability in treated and non-treated keratinocytes can bedetermined using, as a non-limiting example, the In Vitro VascularPermeability assay by Millipore (ECM642). This assay analyzesendothelial cell adsorption, transport, and permeability. Briefly, adulthuman epidermal keratinocytes from Life Technologies (C-005-5C) can beseeded onto a porous collagen-coated membrane within a collection well.The keratinocytes are then incubated for 24 hours at 37° C. and 5% CO₂in Epilife growth media with calcium from Life Technologies(M-EP-500-CA) supplemented with Keratinocyte Growth Supplement (HKGS)from Life Technologies (S-101-5). This incubation time allows the cellsto form a monolayer and occlude the membrane pores. The media is thenreplaced with fresh media with (test sample) or without (non-treatedcontrol) test compounds/extracts and the keratinocytes are incubated foran additional 48 hours at 37° C. and 5% CO₂. To determine permeabilityof the keratinocyte monolayer after incubation with/without the testcompound/extract, the media is replaced with fresh media containing ahigh molecular weight Fluorescein isothiocyanate (FITC)-Dextran and thekeratinocytes are incubated for 4 hours at 37° C. and 5% CO₂. During the4 hours incubation, FITC can pass through the keratinocytes monolayerand porous membrane into the collection well at a rate proportional tothe monolayer's permeability. After the 4 hour incubation, cellviability and the content of FITC in the collection wells can bedetermined. For the FITC content, the media in the collection well iscollected and fluorescence of the media determined at 480 nm (Em) whenexcited at 520 nm. Percent permeability and percent change in comparisonto the non-treated controls can be determined by the followingequations: Percent Permeability=((Mean Ex/Em of test sample)/Mean Ex/Emuntreated control)*100; Percent Change=Percent Permeability of testsample−Percent Permeability of untreated control.

Production of Hyaluronic Acid: Changes in the production of hyaluronicacid in human dermal fibroblasts due to each of the active ingredients,any one of the combination of ingredients, or compositions having saidcombinations disclosed in the specification can be measured. HA is apolysaccharide involved in stabilization of the structure of the matrixand is involved in providing turgor pressure to tissue and cells. As onenon-limiting example, HA production in treated and non-treated adulthuman dermal fibroblasts (HDFa) cells can be determined using theHyaluronan DuoSet ELISA kit from R&D Systems (DY3614). In this assay,for production of samples, subconfluent HDFa cells from CascadeBiologics (C-13-5C) are incubated at 37° C. and 10% CO₂ in starvationmedium (0.15% fetal bovine serum and 1% Penicillin Streptomycin solutionin Dulbecco's Modified Eagle Medium) for 72 hours prior to treatment.The cells are then incubated with fresh starvation medium with eithertest compound, positive control (phorbol 12-myristate 13-acetate fromSigma-Aldrich (P1585) and platelet derived growth factor fromSigma-Aldrich (P3201)), or no additive for 24 hours. Media is thencollected and frozen at −80° C. until use in the ELISA assay.

Briefly, the ELISA assay employs a quantitative sandwich enzymeimmunoassay technique whereby a capture antibody specific for HA can bepre-coated onto a microplate. Standards and media from treated anduntreated cells are pipetted into the microplate wells to enable any HApresent to be bound by the immobilized antibody. After washing away anyunbound substances, an enzyme-linked detection antibody specific for HAis added to the wells. Following a wash to remove any unboundantibody-enzyme reagent, a substrate solution is added to the wells toallow color development in proportion to the amount of HA bound in theinitial step. The color development is stopped at a specific time andthe intensity of the color at 450 nm can be measured using a microplatereader.

Inhibition of Hyaluronidase Activity: Changes in the activity ofhyaluronidase due to each of the active ingredients, any one of thecombination of ingredients, or compositions having said combinationsdisclosed in the specification can be measured. Hyaluronidase is anenzyme that degrades HA. HA is a polysaccharide involved instabilization of the structure of the matrix and is involved inproviding turgor pressure to tissue and cells. As one non-limitingexample, hyaluronidase activity can be determined using an in vitroprotocol modified from Sigma-Aldrich protocol # EC 3.2.1.35. Briefly,hyaluronidase type 1-S from Sigma-Aldrich (H3506) is added to microplatereaction wells containing test compound or controls. Tannic acid can beused as a positive control inhibitor, no test compound can be added forthe control enzyme, and wells with test compound or positive control butwithout hyaluronidase can be used as a background negative control. Thewells are incubated at 37° C. for 10 minutes before addition ofsubstrate (HA). Substrate is added and the reactions incubated at 37° C.for 45 minutes. A portion of each reaction solution is then transferredto and gently mixed in a solution of sodium acetate and acetic acid pH3.75 to stop that portion of the reaction (stopped wells). The stoppedwells and the reaction wells should both contain the same volume ofsolution after addition of the portion of the reaction solution to thestopped wells. Both the reaction wells and the stopped wells areincubated for 10 minutes at room temperature. Absorbance at 600 nm isthen measured for both the reaction wells and the stopped wells.Inhibition can be calculated using the following formulas: Inhibitor (orcontrol) activity=(Inhibitor stopped wells absorbance at 600nm−inhibitor reaction wells absorbance at 600 nm); Initialactivity=control enzyme absorbance at 600 nm; PercentInhibition=[(Initial activity/Inhibitor Activity)*100]−100.

Peroxisome Proliferator-Activated Receptor Gamma (PPAR-γ) Activity:Changes in the activity of PPAR-γ due to each of the active ingredients,any one of the combination of ingredients, or compositions having saidcombinations disclosed in the specification can be measured. PPAR-γ is areceptor critical for the production of sebum. As one non-limitingexample, the activity of PPAR-γ can be determined using a bioassay thatanalyzes the ability of a test compound or composition to inhibitbinding of a ligand. Briefly, fluorescent small-molecule pan-PPARligand, FLUORMONE™ Pan-PPAR Green, available from Life Technologies(PV4894), can be used to determine if test compounds or compositions areable to inhibit binding of the ligand to PPAR-γ. The samples wellsinclude PPAR-γ and fluorescent ligand and either: test compound orcomposition (test); a reference inhibitor, rosiglitazone (positivecontrol); or no test compound (negative control). The wells areincubated for a set period of time to allow the ligand opportunity tobind the PPAR-γ. The fluorescence polarization of each sample well canthen be measured and compared to the negative control well to determinethe percentage of inhibition by the test compound or composition.

Cytokine array: Human epidermal keratinocytes are cultured to 70-80%confluency. The media in the plate is aspirated and 0.025% trypsin/EDTAis added. When the cells became rounded, the culture dish is gentlytapped to release the cells. The trypsin/EDTA containing cells areremoved from the culture dish and neutralized. Cells are centrifuged for5 min. at 180×g to form a pellet of cells. The supernatant is aspirated.The resulting pellet is resuspended in EPILIFE™ media (CascadeBiologics). The cells are seeded in 6-well plates at approximately10-20% confluency. After the cells became approximately 80% confluent,the media is aspirated and 1.0 ml of EpiLife™, along with phorbol13-Myristate 12-acetate (“PMA”) (a known inducer of inflammation) andthe test composition dilutions are added to two replicate wells (i.e.,1.0% (100 μl of 100X stock) and 0.1% (10 μl of 100X stock) testcompositions are diluted into a final volume of 1 ml EpiLife GrowthMedium). The media is gently swirled to ensure adequate mixing. Inaddition, 1.0 ml of EPILIFE™ is added to the control wells, with andwithout additional PMA. The plates are then incubated at 37±1° C. and5.0±1% CO₂ for approximately 5 hours after dosing. Following this 5-hourincubation, all media is collected in conical tubes and frozen at −70°C.

For analysis, a 16-pad hybridization chamber is attached to 16-pad FASTslides arrayed in triplicate with 16 anti-cytokine antibodies plusexperimental controls (Whatman BioSciences), and the slides are placedinto a FASTFrame (4 slides per frame) for processing. Arrays are blockedfor 15 min. at room temp. using 70 ml S&S Protein Array Blocking buffer(Whatman Schleicher and Scheull). Blocking buffer is removed and 70 mlof each supernatant sample is added to each array. Arrays are incubatedfor 3 hours at room temp. with gentle agitation. Arrays are washed 3times with TBS-T. Arrays are treated with 70 ml of an antibody cocktail,containing one biotinylated antibody corresponding to each of thearrayed capture antibodies. Arrays are incubated for 1 hour at roomtemp. with gentle agitation. Arrays are washed 3 times with TBS-T.Arrays are incubated with 70 ml of a solution containingstreptavidin-Cy5 conjugate for 1 hour at room temp. with gentleagitation. Arrays are washed 3 times with TBS-T, quickly rinsed inde-ionized water, and dried.

Slides can be imaged in a Perkin-Elmer ScanArray 4000 confocalfluorescent imaging system. Array images can be saved and analyzed usingImaging Research ArrayVision software. Briefly, spot intensities aredetermined by subtracting background signal. Spot replicates from eachsample condition can be averaged and then compared to the appropriatecontrols.

Endothelial Tube Formation: Endothelial tube formation is involved inangiogenesis and micro-vessel capillary formation. Capillary formationand angiogenesis may contribute to redness and rosacea of the skin. Theability for endothelial cells to form tubes in the presence or absenceof test extracts and compounds may be determined using a capillarytubule disruption assay with pre-formed primary human umbilical veinendothelial cells (HUVEC) in a cell culture system.

Briefly, HUVECs are cultured in vitro on Extracellular Matrix, whichstimulates the attachment and tubular morphogenesis of endothelial cellsto form capillary-like lumen structures. These in vitro formed capillarytubules are similar to human blood vessel capillaries in many aspects.The capillary tube assay is based on this phenomenon and is used forevaluation of potential vasculature targeting agents.

HUVEC cultures are grown in a 5% CO₂ 37° C. cell incubator. The fullgrowth medium for HUVECs is Endothelial Cell Basal Medium (EBM)supplemented with 2% fetal bovine serum (FBS), 12 μg /ml bovine brainextract, 1 μg/ml hydrocortisone, and 1 μg/ml GA-1000(gentamicin-amphothericin). HUVEC cultures between passage 3 and 8 maybe used for all assay experiments.

HUVECs are pre-labeled with fluorescent agent Calcein AM and seeded inExtracellular Matrix coated 96-well culture plate with their full growthmedium. After about four hours of the morphogenesis process, theendothelial capillary tubes should be formed. Then, test agent indesigned doses in 50 μl volume is applied into the formed capillarytubule cultures as treatment conditions. The no-treatment controls canbe added with vehicle of test agents. Sutent, a FDA approvedanti-angiogenic drug one concentration can be included as assayperformance control. After about six hours of treatment, the endothelialtubule morphology in each well is examined by microscopy, imaged, andthe capillary disrupting activities under treatment conditions can bequantitatively analyzed. Each test conditions can be conducted induplicate wells, including controls.

All of the compositions and/or methods disclosed and claimed herein canbe made and executed without undue experimentation in light of thepresent disclosure. While the compositions and methods of this inventionhave been described in terms of preferred embodiments, it will beapparent to those of skill in the art that variations may be applied tothe compositions and/or methods and in the steps or in the sequence ofsteps of the method described herein without departing from the concept,spirit and scope of the invention. More specifically, it will beapparent that certain agents which are both chemically andphysiologically related may be substituted for the agents describedherein while the same or similar results would be achieved. All suchsimilar substitutes and modifications apparent to those skilled in theart are deemed to be within the spirit, scope and concept of theinvention as defined by the appended claims.

REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference.

-   Cosmetic Ingredient Dictionary, Third Edition, CTFA, 1982-   International Cosmetic Ingredient Dictionary, Fourth edition, CTFA,    1991-   International Cosmetic Ingredient Dictionary and Handbook, Tenth    Edition, CTFA, 2004-   International Cosmetic Ingredient Dictionary and Handbook, Twelfth    Edition, CTFA, 2008-   Medications and More During Pregnancy and Breastfeeding, Retinoids;    MotherToBaby;    -   Comprehensive Perinatal Services Program, California Department        of Public Health;    -   https://www.cdph.ca.gov/programs/CPSP/Documents/MotherToBaby.Medication%20and%20More.pdf.        Accessed on May 31, 2016.

1. A method of reducing the appearance of a stretch mark in a person'sskin, the method comprising topically applying to the stretch mark aneffective amount of a topical composition comprising vegetable aminoacids, Myrciaria dubia fruit extract, Peucedanum graveolens extract, andtetrahexyldecyl ascorbate, wherein the appearance of the stretch mark isreduced.
 2. The method of claim 1, wherein the vegetable amino acids arean aqueous extract of navy bean, wherein the Myrciaria dubia fruitextract is an aqueous extract that comprises the pulp of Myrciaria dubiafruit, and wherein the Peucedanum graveolens extract is an aqueousextract.
 3. The method of claim 1, wherein the effective amount of thetopical composition increases collagen density, increases fibrillin-1expression, decreases collagen network orientation, decreases stretchmark visibility, improves the overall appearance of a stretch mark,decreases redness, increases smoothness, increases skin tone/evenness,increases elasticity, increases firmness, improves texture, decreasesthickness of a stretch mark, decreases echogenicity of a stretch mark,inhibits COX-1, MMP1, MMP3, MMP9, and/or tyrosinase activity, increasescollagen production, and/or provides an antioxidant in the skin havingthe stretch mark.
 4. The method of claim 1, wherein the topicalcomposition comprises 0.01 to 5% w/w of vegetable amino acids, 0.01 to5% w/w of Myrciaria dubia fruit extract, 0.01 to 3% w/w of Peucedanumgraveolens extract, and 0.01 to 1% w/w of tetrahexyldecyl ascorbate. 5.The method of claim 1, wherein the topical composition further comprises60 to 85% w/w of water.
 6. The method of claim 1, wherein the topicalcomposition further comprises pentylene glycol, cyclopentasiloxane,glycerin, and hydroxyethyl acrylate/sodium acryloyldimethyl tauratecopolymer.
 7. The method of claim 6, wherein the topical compositionfurther comprises dimethicone, dimethiconol, phenoxyethanol, andethylhexylglycerin.
 8. The method of claim 7, wherein the topicalcomposition further comprises PEG-12 dimethicone, triethanolamine, anddisodium EDTA.
 9. The method of claim 1, wherein the topical compositionis an emulsion, serum, gel, gel emulsion, or gel serum.
 10. The methodof claim 1, wherein the topical composition is applied to a stretch markat least twice a day.
 11. A topical composition comprising vegetableamino acids, Myrciaria dubia fruit extract, Peucedanum graveolensextract, and tetrahexyldecyl ascorbate.
 12. The topical composition ofclaim 11, wherein the vegetable amino acids are an aqueous extract ofnavy bean, wherein the Myrciaria dubia fruit extract is an aqueousextract that comprises the pulp of Myrciaria dubia fruit, and whereinthe Peucedanum graveolens extract is an aqueous extract.
 13. The topicalcomposition of claim 11, wherein the topical composition comprises aneffective amount of vegetable amino acids, Myrciaria dubia fruitextract, Peucedanum graveolens extract, and tetrahexyldecyl ascorbate toincrease collagen density, increase fibrillin-1 expression, decreasecollagen network orientation, decrease stretch mark visibility, improvethe overall appearance of a stretch mark, decrease redness, increasesmoothness, increase skin tone/evenness, increase elasticity, increasefirmness, improve texture, decrease thickness of a stretch mark, and/ordecrease echogenicity of a stretch mark.
 14. The topical composition ofclaim 11, wherein the topical composition comprises an effective amountof Myrciaria dubia fruit extract to inhibit COX-1, MMP1, MMP3, MMP9,and/or tyrosinase activity, increase collagen production, and/or providean antioxidant in skin.
 15. The topical composition of claim 11,comprising 0.1 to 5% w/w of vegetable amino acids, 0.1 to 5% w/w ofMyrciaria dubia fruit extract, 0.1 to 3% w/w of Peucedanum graveolensextract, and 0.01 to 1% w/w of tetrahexyldecyl ascorbate.
 16. Thetopical composition of claim 11, further comprising 60 to 85% w/w ofwater.
 17. The topical composition of claim 11, further comprisingpentylene glycol, cyclopentasiloxane, glycerin, and hydroxyethylacrylate/sodium acryloyldimethyl taurate copolymer.
 18. The topicalcomposition of claim 17, further comprising dimethicone, dimethiconol,phenoxyethanol, and ethylhexylglycerin.
 19. The topical composition ofclaim 18, further comprising PEG-12 dimethicone, triethanolamine, anddisodium EDTA.
 20. The topical composition of claim 11, wherein thetopical composition is an emulsion, serum, gel, gel emulsion, or gelserum.